AU698800B2 - Genetic markers for breast and ovarian cancer - Google Patents

Abstract

Specific BRCA1 mutations, PCR primers and hybridization probes are used in nucleic acid-based methods for diagnostic of inheritable breast cancer susceptibility. Additionally, binding agents, such as antibodies, specific for peptides encoded by the subject BRCA1 mutants are used to identify expression products of diagnostic mutations/rare alleles in patient derived fluid or tissue samples. Compositions with high binding affinity for transcription or translation products of the disclosed BRCA1 mutations and alleles are used in therapeutic intervention. Such products include anti-sense nucleic acids, peptides encoded by the subject nucleic acids, and binding agents such as antibodies, specific for such peptides.

Description

O 96/33271 PCTiUS96/05621 Genetic Markersfor Breast and Ovarian Cancer

INTRODUCTION

Field of the Invention The field of the invention is genetic markers for inheritable breast cancer susceptibility.

Background The largest proportion of inherited breast cancer described so far has been attributed to a genetic locus, the BRCA1 locus, on chromosome 17q21 (Hall et al. 1990 Science 250:1684- 1689; Narod et al. 1991 Lancet 338:82-83; Easton et al. 1993 Am J Hum Genet 52:678-701).

Background material on the genetic markers for breast cancer screening is found in the Jan 29, 1993 issue of Science, vol 259, especially pages 622-625; see also King et al., 1993 J Amer Med Assoc 269:1975-198. Other relevant research papers include King (1992) Nature Genet 2:125- :126; Merette et aL (1992) Amer J Human Genet 50:515-519; NIH/CEPH Collaborative Mapoing Group (1992) Science 258:67-86.

Risks of breast cancer to women inheriting the locus are extremely high, exceeding before age 50 and reaching 80% by age 65 (Newman et al. 1988 Proc Natl Acad Sci USA 85:3044-3048; Hall et al. 1992 Amer J Human Genet 50:1235-1242; Easton et al. 1993).

Epidemiological evidence for inherited susceptibility to ovarian cancer is even stronger (Cramer et al. 1983 J Nati Cancer In.t 71:711-716; Schildkraut Thompson 1988 Amer J Epidemiol 128:456-466; Schildkraut et aL 1989 Amer J Hum Genet 45:521-529). According to one study, more than 90% of families with multiple relatives with breast and ovarian cancer trace disease susceptibility to chromosome 17q21 (Easton et al. 1993).

The link between increasing risk of breast and ovarian cancer and inherited susceptibility Sto these diseases lies in the application of genetics to diagnosis and prevention. Creating molecular tools for earlier diagnosis and developing ways to reverse the first steps of tumorigenesis may be the most effective means of breast and ovarian cancer control.

Our laboratory previously mapped the. heritable breast cancer susceptibility gene locus (BRCA1 locus) to a 50 cM region of chromosome 17q (Hall et al. 1990). More recently, we I. 0 iWH 1 2 1 developed new polymorphisms at ERBB2 (Hall and King 1991 Nucl Acids Res 19:2515), THRA (Bowcock et aL 1993 Amer J Human Genet 52:718-722), EDH17B (Friedman et al. 1993 Hum Molec Genet 2:821), and multiple anonymous loci (Anderson et al. 1993 Genomics 17:6-16-623), ultimately developing a high density map of 17q12-q 2 1 (Anderson et al. 1993; see also, Simard et aL 1993 Human Molec Genet 2:1193-1199). We also added families to the genetic study; there are now 100 families for whom transformed lymphocyte lines have been established and all informative relatives genotyped. We used our new markers and the many chromosome 17q polymorphisms developed in the past three years to test linkage in our families, refining the region first to 8 cM (Hall et al. 1992), then to 4 cM (Bowcock et al. 1993), then to 1 Mb based on polymorphisms from our high density map (Anderson et al. 1993; see also Flejter et al., 1993 Genomics 17:624-631). We disclose here a number of mutations in BRCA1 which correlate with disease.

Relevant Literature The predicted amino acid sequence for a BRCA1 cDNA and familial studies of this gene were described by Miki et al. (1994) Science 266, 66-71 and Futeal et al. (1994) Science 266, .o0 120-122. A study of Canadian cancer families is described in Simard et al. (1994) Nature S Genetics 8, 392-398. A collaborative survey of BRCA1 mutations is described in Shattuch- Eidens et al. (1995) JAMA 273, 535-541.

SUMMARY OF THE INVENTION The invention discloses methods and compositions useful in the diagnosis and treatment of breast and ovarian cancer associated with mutations and/or rare alleles of BRCA1, a breast cancer susceptibility gene. Specific genetic probes diagnostic of inheritable breast cancer susceptibility and methods of use are provided. Labelled nucleic acid probes comprising sequences complementary to specified BRCA1 alleles are hybridized to clinical nucleic acid samples. Linkage analysis and inheritance patterns of the disclosed markers are used to diagnose genetic susceptibility. In addition. BRCA1 mutations and/or rare alleles are directly identified by hybridization, polymorphism and or sequence analysis. I. another embodiment, labeled binding agents, such as antibodies, specific for peptides encoded by the subject nucleic acids are used to identify expression products of diagnostic mutations or alleles in patient derived fluid or tissue samples. For therapeutic intervention, the invention provides comp a:ions which can functionally interfere with the transcription or translation products of the breast and ovarian cancer 1040 *,R4 1, T 0~

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susceptibility associated mutations and/or rare alleles within BRCA1. Such products include antisense nucleic acids, competitive peptides encoded by the subject nucleic acids, and high affinity binding agents such as antibodies, specific for e.g. translation products of the disclosed BRCAI mutations and alleles.

For the purposes of this specification it will be clearly understood that the word "comprising" means "including but not limited to", and that the word "comprises" has a corresponding meaning.

DESCRIPTION OF SPECIFIC EMBODIMENTS We disclose here methods and compositions for determining the presence or absence of BRCA1 mutations and rare alleles or translation products thereof which are useful in the diagnosis of breast and ovarian cancer susceptibility. Tumorigenic BRCA1 alleles include BRCA1 allele #5803 (SEQ ID NO:1), 9601 (SEQ ID NO:2), 9815 (SEQ ID NO:3), 8403 (SEQ ID NO:4), 8203 (SEQ ID NO:5), 388 (SEQ ID NO:6), 6401 (SEQ ID NO:7), 4406 (SEQ ID NO:8), 10201 (SEQ ID NO:9), 7408 (SEQ ID NO:10), 582 (SEQ ID NO:11) or 77 (SEQ ID NO:12). These S nucleic acids or fragments capable of specifically hybridizing with the corresponding allele in the presence of other BRCA1 alleles under stringent conditions find broad diagnostic and therapeutic application. Gene products of the disclosed mutant and/or rare BRCA1 alleles also find a broad :.:range of therapeutic and diagnostic applications. For example, mutant and/or rare allelic BRCA1 peptides are used to generate specific binding compounds. Binding reagents are used diagnostically. to distinguish non-tumorigenic wild-type and tumorigenic BRCA1 translation products.

The subject nucleic acids (including fragments thereof) may be single or double stranded and are isolated, partially purified, and/or recombinant. An "isolated" nucleic acid is present as other than a naturally occurring chromosome or transcript in its natural state and isolated from (not joined in sequence to) at least one nucleotide with which it is normally associated on a natural chromosome; a partially pure nucleic acid constitutes at least about 10%, preferably at least about and more preferably at least about 90% by weight of total nucleic acid present in a given fraction; and a recombinant nucleic acid is joined in sequence to at least one nucleotide with which it is not normally associated on a natural chromosome.

Fragments of the disclosed alleles are sufficiently long for use as specific hybridization 3i *r r WO 96/33271 PCTUS96/0 5 6 2 1 probes for detecting endogenous alleles, and particularly to distinguish the disclosed critical rare Sibl c oe canc from other BRCA1 alleles, including or mutant alleles which correlate with cancer susceptibility from other BRCA1 alleles, including alleles encoding the BRCA1 translation product displayed in Miki et al (1994) supra, under stringent conditions. Preferred fragments are capable of hybridizing to the corresponding mutant allele under stringency conditions characterized by a hybridization buffer comprising 0% formamide in 0.9 M saline/0.

0 9 M sodium citrate (SSC) buffer at a temperature of 37°C and remaining bound when subject to washing at 420C with the SSC buffer at 370C. More preferred fragments will hybridize in a hybridization buffer comprising 20% formamide in 0.9 M saline/0.09 M sodium citrate (SSC) buffer at a temperature of 42°C and remaining bound when subject to washing at 42°C with 2 X SSC buffer at 420C. In any event, the fragments are necessarily of length sufficient to be unique to the corresponding allele; i.e. has a nucleotide sequence at least long enough to define a novel oligonucleotide, usually at least about 14, 16, 18, 20, 22, or 24 bp in length, though such fragment may be joined in sequence to other nucleotides which may be nucleotides which naturally flank the fragment.

In many applications, the nucleic acids are labelled with directly or indirectly detectable signals or means for amplifying a detectable signal. Examples include radiolabels, luminescent fluorescent) tags, components of amplified tags such antigen-labelled antibody, biotin-avidin combinations etc. The nucleic acids can be subject to purification, synthesis, modification, sequencing, recombination, incorporation into a variety of vectors, expression, transfection, administration or methods of use disclosed in standard manuals such as Molecular Cloning,

A

Laboratory Manual (2nd Ed., Sambrook, Fritsch and Maniatis, Cold Spring Harbor), Current Protocols in Molecular Biology (Eds. Aufubel, Brent, Kingston, More, Feidman, Smith and Stuhl, Greene Publ. Assoc., Wiley-Interscience, NY, NY, 1992) or that are otherwise known in the art.

The subject nucleic acids are used in a wide variety of nucleic acid-based diagnostic method that are known to those in the art. Exemplary methods include their use as allele-specific oligonucleotide probes (ASOs), in ligase mediated methods for detecting mutations, as primers in PCR-based methods, direct sequencing methods wherein the clinical BRCA1 nucleic acid sequence is compared with the disclosed mutations and rare alleles, etc. The subject nucleic acids are capable of detecting the presence of a critical mutant or rare BRCA1 allele in a sample and distinguishing the mutant or rare allele from other BRCA1 alleles. For example, where the subject nucleic acids are used as PCR primers or hybridization probes the subject primer or probe 4 re I ip i I ILlqC--~q~~-L -i~Cd-P- Y~-LIL-~I WO 96/33271 PCT/US96/05621 comprises an oligonucleotide complementary to a strand of the mutant or rare allele of length sufficient to selectively hybridize with the mutant or rare allele. Generally, these primers and probes comprise at least 16 bp to 24 bp complementary to the mutant or rare allele and may be as large as is convenient for the hybridizations conditions.

Where the critical mutation is a deletion of wild-type sequence, useful primers/probes require wild-type sequences flanking (both sides) the deletion with at least 2, usually at least 3, more usually at least 4, most usually at least 5 bases. Where the mutation is an insertion or substitution which exceeds about 20 bp, it is generally not necessary to include wild-type sequence in the probes/primers. For insertions or substitutions of fewer than 5 bp, preferred nucleic acid portions comprise and flank the substitution/insertion with at least 2, preferably at least 3, more preferably at least 4, most preferably at least 5 bases. For substitutions or insertions from about 5 to about 20 bp, it is usually necessary to include both the entire insertion/substitution and at least 2, usually at least 3, more usually at least 4, most usually at least 5 basis of wild-type sequence of at least one flank of the substitution/insertion.

In addition to their use as diagnostic genetic probes and primers, BRCA1 nucleic acids are used to effect a variety of gene-based therapies. See, e.g. Zhu et al. (1993) Science 261, 209-211; Gutierrez et al. (1992) Lancet 339, 715-721; Gary Nabel lab (Dec 1993), Proc. Nat'l. Acad Sci USA. For example, therapeutic nucleic acids are used to modulate cellular expression or intracellular concentration or availability of a tumorigenic BRCA1 translation product by introducing into cells complements of the disclosed nucleic acids. These nucleic acids are typically antisense: single-stranded sequences comprising complements of the disclosed relevant BRCA1 mutant. Antisense modulation of the expression of a given mutant may employ antisense nucleic acids operably linked to gene regulatory sequences. Cell are transfected with a vector comprising such a sequence with a promoter sequence oriented such that transcription of the gene yields an antisense transcript capable of binding to the endogenous tumorigenic BRCA 1 allele or transcript. Transcription of the antisense nucleic acid may be constitutive or indu .ble and the vector may provide for stable extrachromosomal maintenance or integration. Alternatively, single-stranded antisense nucleic acids that bind to BRCA1 genomic DNA or mRNA may be administered to the target cell, in or temporarily isolated from a host, at a concentration that results in a substantial reduction in expression of the targeted translation product.

Various techniques may be employed for introducing of the nucleic acids into viable cells.

WO 96/33271 PCT/US96/0562 1 The techniques vary depending upon whether one is using the subject compositions in culture or in vivo in a host. Various techniques which have been found efficient include transfection with a retrovirus, viral coat protein-liposome mediated transfection, see Dzau et al., Trends in Biotech 11, 205-210 (1993). In some situations it is desirable to provide the nucleic acid source with an agent which targets the target cells, such as an antibody specific for a surface membrane protein on the target cell a ligand for a receptor on the target cell, etc. Where liposomes are employed, proteins which bind to a surface membrane protein associated with endocytosis may be used for targeting and/or to facilitate uptake, e.g. capsid proteins or fragments thereof tropic for a particular cell type, antibodies for proteins which undergo internalization in cycling, proteins that target intracellular localization and enhance intracellular half-life. In liposomes, the decoy concentration in the lumen will generally be in the range of about 0.1 [M to 20 pM. For other techniques, the application rate is determined empirically, using conventional techniques to determine desired ranges. Usually, application of the subject therapeutics will be local, so as to be administered at the site of interest. Various techniques can be used for providing the subject compositions at the site of interest, such as injection, use of catheters, trocars, projectiles, pluronic gel, stents, sustained drug release polymers or other device which provides for internal access.

Systemic administration of the nucleic acid using lipofection, liposomes with tissue targeting (e.g.

antibody) may also be employed.

The invention also provides isolated translation products of the disclosed BRCA1 allele which distinguish the wild type BRCA1 gene product. For example, for alleles which encode truncated tumorigenic translation product, the C-terminus is used to differentiate wild-type BRCA1. Accordingly, the invention provides tle translation product of BRCA1 allele #5803 (SEQ ID NO:13), 9601 (SEQ ID NO:14), 9815 (SEQ ID NO:15), 8203 (SEQ ID NO:17), 388 (SEQ ID NO:18), 6401 (SEQ ID NO:19), 4406 (SEQ ID NO:20), 10201 (SEQ ID NO:21), 7408 (SEQ ID NO:22), 582 (SEQ ID NO:23) or 77 (SEQ ID NO:24), or a C-terminus fragment thereof; and that of #8403 (SEQ ID NO:16), or a fragment thereof cor ising Gly at position 61.

The subject mutant and/or rare allelic BRCA1 translation products comprise an amino acid sequence which provides a target for distinguishing the product from that of other BRCA1 alleles.

Preferred fragments are capable of eliciting the production of a peptide-specific antibody, in vivo or in vitro, capable of distinguishing a protein comprising the immunogenic peptide from a wildtype BRCA1 translation product. The fragments are necessarily unique to the disclosed allele 6 i WO 96/33271 PCTIUS96/05621 translation product in that it is not found in any previously known protein and has a length at least long enough to define a novel peptide, from about 5 to about 25 residues, preferably from 6 to residues in length, depending on the particular amino acid sequence.

The subject translation products (including fragments) are either isolated, i.e.

unaccompanied by at least some of the material with which they are associated in their natural state); partially purified, i.e. constituting at least about preferably at least about 10%, and more preferably at least about 50% by weight of the total translation product in a given sample; or pue, i.e. at least about 60%, preferably at least 80%, and more preferably at least about by weight of total translation product. Included in the subject translation product weight are any atoms, molecules, groups, etc. covalently coupled to the subject translation products, such as detectable labels, glycosylations, phosphorylations, etc. The subject translation products may be isolated, purified, modified or joined to other compounds in a variety of ways known to those skilled in the art depending on what other components are present in the sample and to what, if anything, the translation product is covalently linked.

Binding agents specific for the disclosed tumorigenic BRCA1 genes and gene products find particular use in cancer diagnosis. The selected method of diagnosis will depend on the nature of the tumorigenic BRCA1 mutants/rare allele and its transcription or translation product(s). For example, soluble secreted translation products of the disclosed alleles may be detected in a variety of physiologic fluids using a binding agent with a detectable label such as a radiolabel, fluorescer etc. Detection of membrane bound or intracellular products generally requires preliminary isolation of cells blood cells) or tissue breast biopsy tissue). A wide variety of specific binding assays, e.g. ELISA, may be used BRCA1 gene product-specific binding agents are produced in a variety of ways using the compositions disclosed herein. For example, structural x-ray crystallographic and/or NMR data of the mutant and/or rare allelic BRCA1 translation products are used to rationally d ign binding molecules of determined structure or complementarity. Also, the disclosed mutant and/or rare allelic BRCA1 translation products are used as immunogens to generate specific polyclonal or monoclonal antibodies. See, Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, for general methods. Specific antibodies are readily modified to a monovalent form, such as Fab, Fab', or Fv.

Other mutant and/or rare allelic BRCA1 gene-product specific agents are screened from 7 I L t I WO 96/33271 PCT/US96/05621 large libraries of synthetic or natural compounds. For example, numerous means are available for random and directed synthesis of saccharide, peptide, and nucleic acid based compounds.

Alternatively, libraries of natural compounds in the form of bacterial, fungal, plant and animal extracts are available or readily producible. Additionally, natural and synthetically produced libraries and compounds are readily modified through conventional chemical, physical, and biochemical means. See, e.g. Houghten et al. and Lam et al (1991) Nature 354, 84 and 81, respectively and Blake and Litzi-Davis (1992), Bioconugate Chem 3, 510.

Useful binding agents are identified with assays employing a compound comprising mutant and/or rare allelic BRCA1 peptides or encoding nucleic acids. A wide variety of in vitro, cell-free binding assays, especially assays for specific binding to immobilized compounds comprising the subject nucleic acid or translation product find convenient use. See, e.g. Fodor et al (1991) Science 251, 767 for the light directed parallel synthesis method. Such assays are amenable to scale-up, high throughput usage suitable for volume drug screening.

Useful agents are typically those that bind the targeted mutant and/or rare allelic BRCA1 gene product with high affinity and specificity and distinguish the tumorigenic BRCA1 mutants/rare alleles from the wild-type BRCA1 gene product. Candidate agents comprise functional chemical groups necessary for structural interactions with proteins and/or DNA, and typically include at least an amine, carbonyl, hydroxyl or carboxyl group, preferably at least two of the functional chemical groups, more preferably at least three. The candidate agents often comprise cyclical carbon or heterocyclic structures and/or aromatic or polyaromatic structures substituted with one or more of the forementioned functional groups. Candidate agents are also found among biomolecules including peptides, saccharides, fatty acids, sterols, isoprenoids, purines, pyrimidines, derivatives, structural analogs or combinations thereof, and the like. Where the agent is or is encoded by a transfected nucleic acid, said nucleic acid is typically DNA or

RNA.

Candidate agents are obtained from a wide variety of sources including libraries of synthetic or natural compounds. For example, numerous means are available for random and directed synthesis of a wide variety of organic compounds and biomolecules, including expression of randomized oligonucleotides. Alternatively, libraries of natural compounds in the form of bacterial, fungal, plant and animal extracts are available or readily produced. Additionally, natural and synthetically produced "'raries and compounds are readily modified through conventional rce i- i_ :4 1IL-~ pc-il.- y-l -Y ~1II WO 9633271 CTUS96/05621 WO 96/33271 chemical, physical, and biochemical means to enhance efficacy, stability, pharmaceutical compatibility, and the like. In addition, known pharmacological agents may be subject to directed or random chemical modifications, such as acylation, aE, i tion, esterification, amidification, etc., to produce structural analogs.

Therapeutic applications typically involve binding to and functional disruption of a tumorigenic BRCA1 gene product by an administered high affinity binding agent. For therapeutic uses, the compositions jnd agents disclosed herein may be administered by any convenient way.

Small organics are preferably administered orally; other compositions and agents are preferably administered parenterally, conveniently in a pharmaceutically or physiologically acceptable carrier, phosphate buffered saline, or the like. Typically, the compositions are added to a retained physiological fluid such as blood or synovial fluid. Generally, the amount administered will be empirically determined, typically in the range of about 10 to 1000 pg/kg of the recipient. For peptide agents, the concentration will generally be in the range of about 50 to 500 pg/ml in the dose administered. Other additives may be included, such as stabilizers, bactericides, etc. These additives will be present in conventional amounts.

The following examples are offered by way of illustration and not by way of limitation.

EXAMPLES

Example 1. Positional cloning Contig construction.

YACs. Primers flanking polymorphic repeats in the 4 Mb region of linkage were used to amplify pools from the CEPH, Washington University, and CEPH megaYAC libraries available. 39 YACs were selected. Of these, 23 were tested for chimerism by FISH and 12 found to be chimeric.

YACs were aligned to each other by attempting to amplify each YAC with primer pairs from known sequence tagged sites (STSes). More STSes were defined by sequencing the ends of YACs, and these new STSes used ir further alignment and YAC identification.

Cosmids. A gridded cosmid library of chromosome 17 was prepared. Alu-Alu PCR products of YACs were hybridized to the cosmid grids and positively hybridizing cosmids used for subsequent studies. Contigs were constructed in two ways. Cosmids with the same restriction patterns were aligned; and, the unique sequences flanking polymorphic markers and our sequenced cDNAs were used as STSes.

9 L' WO 96/33271 PCT/US96/05621 Physical mapping by pulsed fiela gel electrophoresis. Physical distances were estimated by pulsed field gel electrophoresis, using DNA from lymphocyte cell lines of BRCAl-linked patients and of controls. DNA samples were digested with NotI, Mlul, RsrII, NruI, SacII, and EclXI. Filters were probed with single-copy sequences isolated from cosmids and later with cDNA clones. Multiple unrelated linked patients and controls were screened to detect large insertions or deletions associated with BRCA1. Results of PFGE were used to define the region first used to screen cDNA libraries as 1 Mb and the current linked region as 500 kb.

Screening cDNA libraries. We began library screening when the linked region defined by meiotic recombination was -1 Mb. The first question was what library would optimize the length of cDNA clones, representation of both 5' and 3' ends of genes, and the chances that BRCA1 would be expressed. We chose to use a random primed cDNA library cloned into IgtlO from cultured (not transformed) fibroblasts from a human female. This library was selected because it had inserts averaging 1.8 kb, with 80% of inserts between 1 and 4 kb, was contructed from cultured fibroblasts known to be "leaky" in gene expression, and was known to include 5' ends of genes. We simultaneously screened three other libraries (from ovary, fetal brain, and mouse mammary epithelium). With one exception (described below), all tr-- ripts from these libraries cross-hybridized to transcripts from the fibroblast library.

The fibroblast library was screened with YAC DNA isolated by PFGE. Pure YAC DNA (100 nanograms) was random primed with both aP32-dATP (6000mCi/mmole) and 32 P-dCTP (3000 mCi/mmole), and used immediately after labelling. Filters from the library were prehybridized with human placental DNA for 24-48 hours. Labelled YAC DNA was hybridized to the filters for 48 hours at 65C. Approximately 250 transcripts were selected by screening with 7 YACs and then ross-hybridized. We also used pools of cosmids from the linked region to screen the fibroblast library. We selected 122 transcripts and cross-hybridized them to clones previously detected by the YACs.

Example 2. Cloning BRCA1 and its characterization.

A. Screening for mutations in candidate genes. We initially identified 24 genes in the 1Mb BRCA1 region defined by meiotic recombinatio,, respective locations on the YAC coltig, sizes of representative cDNA clones, numbers of rn cates in the library, sizes of transcripts, homologies to known genes, and variants detected. Candidate gene were characterized in the following ways: WO 96/33271 PCTIUS96/05621 Cross-hybridizing clones. cDNA clones isolated from the library are hybridized against each other. Cross-hybridizing clones are considered "siblings" of the clone used as a probe and represent the same gene.

Mapping back. At least one clone from each sibship is mapped back to total human genomic DNA, to cosmids, to YACs, and to somatic cell hybrid lines, some of which contain deletions of 17q and one of which has chrorrosome 17 as its only human chromosome.

Subcloning andseuencing. One of the longest clones from each sibship is subcloned into M13 and sequenced manually by standard methods, constructing new primers at the end of each fragment to continue sequencing until the end of the clone is reached.

Extnding s nces with sibs. In order to find clones that contain more of the gene, the last sequencing primer for the clone and primers made from Xgtl0 are used to amplify sibs of the first clone. Sibs that amplify the longest fragments are selected, subclored, and sequenced. This process is continued until we rea-h the size of the transcript defined by Northern blot and/or until the 3' sequence is a polyA tail and the 5' sequence has features of the beginning of the coding region.

Southerns. To identify insertion or deletion mutations, ge. DNA from 20 unrelated patienb from families with breast cancer linked to 17q "linked patients") and controls are digested with BamI/TaqI and independently with HindII/HinfI. Each cDNA clone is ued to screen Southern blots. Variants have been detected in two genes. Both of these variants are RFLPs, occuring in equal frequency in linked patients and in controls.

NoQthrns. To identify splice mutations and/or length mutations, we prepared total RNA and polyA+ RNA from germline DNA (from lymphoblast lines) of 20 unrelated linked patients, from ovarian and breast tissues, from fibroblasts, from a HeLa cell line, and from breast cancer cell lines. Northern blots are screened with each gene.

Detection of small mutations. To screen for g.rmline point mutations in patients without encountering introns, prepared cDNA from poly-A+ mRNA from lymphoblast cell lines of unrelated linked patients and from controls. cDNA has also been made from 65 malignant ovarian cancers from patients not selected for family history. Primers are constructed every -200 basepairs along the sequence and used to amplify these cDNAs. Genomic DNA has also been prepared from cell lines from all family members (linked and unlinked), from malignant and normal cells from paraffin blocks from their bieast and ovarian surgeries, and from malignant and normal 11 i

I

WO 96/33271 PCT/US96/05621 cells from 29 breast tumors not selected for family history. For sequences without introns, cDNA and gDNA lengths are equal, and the gDNA samples are amplified as well.

Two mutation detection methods are used to screen each sequence. Amplified products are screened for SSCPs using modifications that enable electrophoresis to be done with only one set of running conditions (Keen et al. 1991 Trends Genet 7:5; Soto and Sukumar 1992 PCR Meth Appl 2:96-98). In order to screen longer segments of DNA (100-1500 bp) and to detect variants missed by SSCP, sequences are also screened for point mutations by CCM (Cotton 1993 Mutation Res 285:125-144) using essentially the protocol of Grompe et al. 1989 Proc Natl Acad Sci USA 86:5888-5892. An endonuclease developed for mismatch detection reduces the toxicity of the method (Youil et al. 1993 Amer J Hum Genet 53 (supplement): abstract 1257).

Polymorphism or mutation. Variants are screened in cases and controls to distinguish polymorphisms from a critical mutation. Linkage of breast cancer to each variant is tested in all informative families.

Example 3. Characterize BRCA1 mutations in germline DNA and breast cancer patients tumors.

A. BRCA1 mutations inchromosome 17q-linked families. Our seiies of families includes large extended kindreds in which breast and ovarian cancer (and in one family prostatic cancer) are linked to 17q21, with individual lod scores 1.5. Since linked patients in these i milies carry mutations in BRCA1, we have identified their mutations first.

Table 1 summarizes critical BRCA1 mutations and rare alleles: Family Exon U14680 nt Mutation Amino Acid Predicted __change effect 5803 3 200-253 exon 3 deleted (54 bp) 27 Stop protein truncation 9601 3 230 deletion AA 39 Stop protein truncation 9815 Intron 5 splice donor, substitution G to A 64 Stop protein bp +1 ->22 bp deletion in RNA truncation 8403 5 300 substitution T to G Cys 61 Gly lose zincbinding motif 8203 Intron 5 splice substitution T to G 81 Stop protein acceptor, bp ->59 bp insertion of truncation -11 intron into RNA i 1 WO 9 WgP 96/33271 PCT/S96/0562 388 11 1048 deletion A 313 Stop protein truncation 6401 11 2415 deletion AG Ser 766 Stop protein truncation 4406 11 2800 deleiton AA 901 Stop protein truncation 10201 11 2863 deletion TC Ser 915 Stop protein truncation 7408 11 3726 substitution C to T Arg 1203 protein Stop truncation 582 11 418, deletion TCAA 1364 Stop protein truncation 77 24 5677 Insertion A Tyr 1853 protein Stop truncation

U

B. Germline BRCA I mutations among breast cancer patients in the general population.

From each breast cancer patient, not selected for family history, a 30 ml sample of whole blood is drawn into acid citrate dextrose. DNA from the blood is extracted and stored at in 3 aliquots. Germline mutations in BRCA1 are identified using the approaches described above and by directly sequencing new mutations. Paraffin-embedded tumor specimens from the same patients are screened for alterations of p53, HER2, PRAD1, and ER. Germline BRCA1 mutations are tested in the tumor blocks.

A preliminary estimate of risk associated with different BRCA1 mutations is obtained from relatives of patients with germline alterations. For each patient with a germline BRCA1 mutation, each surviving sister and mother (and for older patients, brothers as well), DNA is extracted from a blood sample and tested for the presence of the proband's BRCA1 mutation. To ascertain men at risk of prostatic cancer, brothers of breast cancer patients diagnosed after age 55 are also interviewed and sampled. Paraffin blocks from deceased relatives who had cancer are also screened. The frequency of breast, ovarian, or prostatic cancer among relatives carrying BRCA 1 mutations is a first estimate of risk of these cancers associated with different mutations. Somatic alterations of BRCA1 in breast tumors.

Malignant cells are dissected from normal cells from paraffin blocks. By identifying BRCA1 mutations in these series, we estimate the frequency of somatic BRCA1 alterations, determine BRCA1 mutations characteristic of any particular stage of tumor development, and 13 (i*y WO 96/33271 PCT/US96/05621 evaluate their association with prognosis.

D. Characterizing mutant and rare alleles of BRCA1. Mutant or rare BRCA1 allele function and pattern of expression during development are characterized using transformed cells expressing the allele and knockout or transgenic mice. For example, phenotypic changes in the animal or cell line, such as growth rate and anchorage independence are determined. In addition, several methods are used to study loss-of-function mutations, including replacing normal genes with their mutant alleles (BRCA1-/BRCA1-) by homologous recombination in embryonic stem (ES) cells and replacing mutant alleles with their normal counterparts in differentiated cultured cells (Capecchi 1989 Science 244:1288-1292; Weissman et al. 1987 Science 236:175-180; Wang et al. 1993 Oncogene 8:279-288). Breast carcinoma cell lines are screened for mutation at the BRCA1 locus and a mutant BRCA1 line is selected. Normal and mutant cDNAs of BRCA1 are subcloned into an expression vector carrying genes which confer resistance to ampicillin and geneticin (Baker et al. 1990 Nature 249:912-915). Subclones are transfected into mutant BRCA1 breast cancer cells Geneticin-resistant colonies are isolated and examined for any change in tumorigenic phenotype, such as colony formation in soft agar, increased growth rate, and/or tumor formation in athymic nude mice. In vivo functional demonstrations involve introducing the normal BCRA1 gene into a breast carcinoma cell line mutant at BRCA1 and injecting these BRCAl+ cells into nude mice. Changes observed in tumorigenic growth compared to nude mice injected with BRCA1 mutant breast carcinoma cells are readily observed. For example, correcting the mutant gene decreases the ability of the breast carcinoma cells to form tumors in nude mice (Weissman et al. 1987; Wang et al. 1993).

All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and cxam-.e for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

14 S14 1 H ryl -3 I T 0 WO 96/33271 PCTIUS96/05621 SEQUENCE LISTING GENERAL INFORMATION: APPLICANT: KING, Mary-Claire FRIEDMAN, Lori OSTERMEYER, Beth ROWELL, Sarah LYNCH, Eric SZABO, Csilla LEE, Ming (ii) TITLE OF INV7NTION: GENETIC MARKERS FOR BREAST AND OVARIAN

CANCER

(iii) NUMBER OF SEQUENCES: 24 (iv) CORRESPONDENCE ADDRESS: ADDRESSEE: Science Technology Law Group STREET: 268 Bush Street, Suite 3200 CITY: San Francisco STATE: California COUNTRY: USA ZIP: 94104 COMPUTER READABLE FORM: MEDIUM TYPE: Floppy disk COMPUTER: IBM PC compatible OPERATING SYSTEM: PC-DOS/MS-DOS SOFTWARE: PatentIn Release Version #1.30 (vi) CURRENT APPLICATION DATA: APPLICATION NUMBER: US FILING DATE:

CLASSIFICATION:

(viii) ATTORNEY/AGENT INFORMATION: NAME: OSMAN, Richard A REGISTRATION NUMBER: 36,627 REFERENCE/DOCKET NUMBER: A-59563-3/RAO (ix) TELECOMMUNICATION INFORMATION: TELEPHONE: (415) 343-4341 TELEFAX: (415) 343-4342

TELEX:

INFORMATION FOR SEQ ID NO:1: SEQUENCE CHARACTERISTICS: LENGTH: 5656 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG GTTTCTCAGA TAACTGGGCC CCTGCGCTCA GGAGGCCTTC ACCCTCTGCT CTGGGTAAAG TTCATTGGAA CAGAAAGAAA 120 TGGATTTATC TGCTCTTCGC GTTGAAGAAG TACAAAATGT CATTAATGCT ATGCAGAAAA 180 TCTTAGAGTG TCCCATCTGA TTTTGCATGC TGAAACTTCT CAACCAGAAG AAAGGGCCTT 240 CACAGTGTCC TTTATGTAAG AATGATATAA CCAAAAGGAG CCTACAAGAA AGTACGAGAT 300 TTAGTCAACT TGTTGAAGAG CTATTGAAAA TCATTTGTGC TTTTCAGCTT GACACAGGTT 36C TGGAGTATGC AAACAGCTAT AATTTTGCAA AAAAGGAAAA TAACTCTCCT GAACATCTAA 420 AAGATGAAGT TTCTATCATC CAAAGTATGG GCTACAGAAA CCGTGCCAAA AGACTTCTAC 480 I ;Mmm WO 96/33271 PCTJUS96OS621

AGAGTGAACC

TTGGAACTGT

ACATTGAATT

TGGGAGATCA

ATTCTGCAAA

ATCAACCCAG

AAAAGTATCA

ATGCCAGCTC

TAGAAAAGGC

ACAGATGGGC

AGGTAGATCT

CATGCTCAGA

TTCAGAAAGT

ATGATGGGGA

TAGATGAATA

CTTTAATATG

AAATATTTGG

AAAATCTAAT

CAAATAAATT

AGAAAGCAGA

CGGAGCAGI~k

GTGATTCTAT

CTTTCAAAAC

ATATCCACAA

ATATTCATGC

TGCAAATTGA

CAGTCAGGCA

AGAAGAGTAA

AGCTGAAGrT

AAGAATTTGT

AAGTGTCTAA

AAACTGAAAG

CGAAAATCCT

GAGAACTCTG

GGGATCTGAT

AGAATTGTTA

AAAGGCTGCT

TAATAATGAT

GGGTAGTTCT

ATTACAGCAT

TGAAT'rCTGT

TGGAAGTAAG

GAATGCTGAT

GAATCCTAGA

TAATG. jTGG

GTCTGAATCA

TTCTGGTTCT

TAAAAGTGAA

GAAAACCTAT

TATAGGAGCA

AA.AGCGTAAA

TTTGGCAGTT

TGGTCAAGTG

TCAGAATGAG

GAAAGCTGAA

TTCAAAAGCA

GCTTGAACTA

TAGTTGTTCT

CAGCAGAA

C;UXGCCAAAT

AACAAATGCA

CAATCCTAGC

TAATGCTGAA

ATCTGTAGAG

TCCTTGCAGG

AGGACAAAGC

TCTTCTGAAG

CAAATCACCC

TGTGAATTTT

TTGAACACCA

GTTTCAAACT

GAGAACAGCA

AATAAAGCA

GAAACATGTA

CCCCTGTGTG

GATACTGAAG

TTTTCCAGAA

AATGCCAAAG

TCAGAGAAAA

AGAGTTCACT

CGGAAGAAGG

TTTGTTACTG

.AGGAGACCTA

CAAAAGACTC

ATGAATATTA

AAAAATCCTA

CCTATAAGCA

CCTAAAAAGA

GTAGTCAGTA

AGCAGTGAAG

CTACAACTCA

GAACAGACAA

CCTGGTTCTT

CTTCCAAGAG

GACCCCAAAG

AGTAGCAGTA

AAACCAGTCT CAGTGTCCAA AGCGGATACA ACCTCAAAAG ATACCGTTAA TAAGGCAACT i

CTCAAGGAAC

CTGAGACGGA

CTGAGAAGCG

TGCATGTGGA

GTTTATTACT

AACAGCCTGG

ATGATAGGCG

AGAGAAAAGA

ATGTTCCTTG

GTGATGAACT

TAGCTGATGT

TAGACTTACT

CCAAATCAGT

CAAGCCTCCC

AGCCACAGAT

CATCAGGCCT

CTGAAATGAT

CTAATAGTGG

ACCCAATAGA

GCAGTAkTAAG

ATAGGCTGAG

GAAATCTAAG

AGATAAAGAA

TGGAAGGTAA

GTAAAAGACA

TTACTAAGTG

AAGAAAAAGA

ATCTCATGTT

TTTCATTGGT

CAGGGATGAA

TGTAACAAAT

TGCAGCTGAG

GCCATGTGGC

CACTAAAGAC

CTTAGCAAGG

GACTCCCAGC

ATGGAATAAG

GATAACACTA

GTTAGGTTCT

ATTGGACGTT

GGCCAGTGAT

AGAGAGTAAT

CAACTTAAGC

AATACAAGAG

TCATCCTGAG

AAATCAGGGA

TCATGAGAAT

ATCACTCGAA

CAATATGGAA

GAGGAAGTCT

CCCACCTAAT

AAAAAAGTAC

AGAACCTGCA

TGACAGCGAT

TTCAAATACC

AGAGAAACTA

AAGTGGAGAA

ACCTGGTACT

CTCTCTAACC

ACGTCTGTCT

TATTGCAGTG

ATCAGTTTGG

ACTGAACATC

AGGCATCCAG

ACAAATACTC

AGAATGAATG

AGCCAACATA

ACAGAAAAAA

CAGAAACTGC

AATAGCAGCA

GATGACTCAC

CTAAATGAGG

CCTCATGAGG

ATTGAAGACA

CATGTAACTG

CGTCCCCTCA

GATTTTATCA

ACTAACCAAA

AAAACAAAAG

AAAGAATCTG

CTCGAATTAA

TCTACCAGGC

TGTACTGAAT

AACCAAATGC

ACTGGAGCCA

ACTTTCCCAG

AGTGAACTTA

GAAACAGTTA

AGGGTTTTGC

GATTATGGCA

540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 WO 96133271 WO 9633271PCTIUS96/05621 CTCAGGAAA:3 TATCTCGTrTA ATAAATGTGT GAGTCAGTGT CCAAAGATAA TAGAAATGAC ACAGTCGGGA AACAAGCATA ATACATTCAA GGTTTCAAAG AAGAGGAATG TGCAACATTC TCACTTTTGA ATGTGAACAA CTGTACAGAC AGTTAATATC TTGATAATGC CAAATGTAGT GAGGCAACGA AACTGGACTC GTATACCACC ACTTTTTCCC TAGAGGAAAA CTTTGAGGAA TTCCAAGTAC AGTGAGCACA

CCAGCTCAAG

ATGAAATAGG

AATTGAATGC

CTGGAAGTA

CTGTTAATAC

GTAGTCATGC

TAAAGGAAGA

AAAGCGTCCA

CTCAGGGTTA

AGGATGAAGA

CTCAGTCTAC

ATTTATTATC

CATCTCAGGA

AGTGCAGTGA

GTTCTTCCAA

AATTGGTTTC

AAAGCATGGA

CAATATTAAT

TTCCAGTGAT

TATGCTTAGA

TTGTA.XGCAT

AGATTTCTCT

ATCTCAGGTT

TACTAGTTTT

GAAAGGAGAG

CCGAAGAGGG

GCTTCCCTGC

TAGGCATAGC

ATTGAAGAAT

ACATCACCTT

ATTGGAAGAC

ACAAATGAGG

AGATGATGAA

TTCAAACTTA

CTGGAAGTTA

GCAGCATTTG

ACAGAAGGCT

GAAATGGAAG

CGCCAGTCAT

TCTGCCCACT

AAGGAAGAAA

ACTGCAGGCT

ATCAAAGGAG

ATTACTCCAA

ATCAAGTCAT

CATTCAATGT

ATTAGCCGTA

GAAGTAGGTT

GAAAACATTC

TTAGGGGTTT

CCTGAAATAA

CCATATCTGA

TGTTCTGAGA

GCTGAAAATG

CTTAGCAGGA

GCCAAGAAAT

TTCCAACACT

ACCGTTGCTA

AGCTTAAATG

AGTGAGGAAA

TTGACTGCAA

CATCAGTCTG

GAAAGAGGAA

GGTGAAGCAG

GCACTCTAGG

AAAkACCCCAA

TTAAGTATCC

AAAGTGAACT

TTGCTCCGTT

CTGGGTCCTT

ATCAAGGAAA

TTCCTGTGGT

GCTCTAGGTT

ATAAACATGG

TTGTTAAA.AC

CACCTGAAAG

ATAACATTAG

CCAGTACTAA

AAGCAGAACT

TGCAACCTGA

AAAAGCAAGA

TTTCAGATAA

CACCTGATGA

ACATTAAGGA

GTCCTAGCCC

TAGAGTCCTC

TGTTATTTGG

CCGAGTGTCT

ACTGCAGTAA

CAAAATGTTC

ATACAAACAC

AA.AGCCAGGG

CGGGCTTGGA

CATCTGGGTG

GAAGGCAAAA

GGGACTAATT

ATTGGGACAT

TGATGCTCAG

TTCAAATCCA

AAAGAAACAA

GAATGAGTCT

TGGTCAGAA.A

TTGTCTATCA

ACTTTTACAA

TAAAkTGTAAG

AGAAATGGGA

AGAA.AATGTT

TGAAGTGGGC

AGGTAGAAAC

GGTCTATAAA

ATATGAAGAA

CTTAGAACAG

CCTGTTAGAT

AAGTTCTGCT

TTTCACCCAT

AGAAGAGAAC

TAAAGTAAAC

GTCTAAGAAC

CCAGGTAATA

TGCTAGCTTG

CCAGGATCCT

AGTTGGTCTG

AGAZAAAAT

TGAGAGTGAA

AACCACTCAG

TGAACTAGAA

ACAGAACCAA

CATGGTTGTT

GAAGTTAACC

TATTTGCAGA

GGAAATGCAG

AGTCCAAAAG

AATATCA.AGC

GATAAGCCAG

TCTCAGTTCA

AACCCATATC

AAAAATCTGC

AATGAGAACA

TTTAAAGAAG

TCCAGTATTA

AGAGGGCCAA

C-AAGTCTTC

GTAG-TTCAGA

CCTATGGGAA

GATGGTGAAA

GTTTTTAGCA

ACACATTTGG

TTATCTAGTG

AATATACCTT

AC-IGAGGAGA

TTGGCAAAGG

TTTTCTTCAC

TTCTTGATTG

AGTGACAAGG

CAAGAAGAGC

ACAAGCGTCT

CAGAGGGATA

GCTGTGTTAG

2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 CCATGCAACA TAACCTGATA AAGCTCCAGC AGGA-AATGG 17

T

c WO 96/33271

AACAGCATGG

TTGAGGACCT

AAAGTAGTGA

TGTCTGCAGA

CTAAATGCCC

ATAGAAACTA

TGGAAGAGTC

AGGGAACCCC

CTTCTGAAGA

CATTGAAAGT

CTACTGATAC

TGACAGCTTC

CAGAAGAATT

TAATTACTGA

GGACACTGAA

TGACCCAGTC

ATGTGGTCAA

AGATCTTCAG

AACTGGAATG

CCCTTGGCAC

ATGGCTTCCA

TGGACAGTGT

GAGCCAGCCT

GCGAAATCCA

ATACCCTATA

TAGTTCTACC

ATCATMAGAT

CCCATCTCAA

TGGGCCACAC

TTACCTGGAA

CAGAGCCCCA

TCCCCAATTG

TGCTGGGTAT

AACAGAAAGG

TATGCTCGTG

AGAGACTACT

ATATTTTCTA

TATTAAAGAA

TGGAAGAAAC

GGGGCTAGAA

GATGGTACAG

AGGTGTCCAC

TGCAATTGGG

AGCACTCTAC

TCTAACAGCT

GAACAAAGCA

AGCCAGAATC

AGTAAAAATA

GATAGGTGGT

GAGGAGCTCA

GATTTGACGG

TCTGGAATCA

GAGTCAGCTC

AAAGTTGCAG

AATGCAATGG

GTCAACAAAA

TACAAGTTTG

CATGTTGTTA

GGAATTGCGG

AGAAAAATGC

CACCAAGGTC

ATCTGTTIGCT

CTGTGTGGTG

CCAATTGTGG

CAGATGTGTG

CAGTGCCAGG

ACCCTTCCAT

CATCAGAAAA

CAGAAGGCCT

AAGAACCAGG

ACATGCACAG

TTAAGGTTGT

AAACATCTTA

GCCTCTTCTC

GTGTTGGCAA

AATCTGCCCA

AAGAAAGTGT

GAATGTCCAT

CCAGAAAACA

TGAAAACAGA

GAGGAAAATr TGAATG.Z1

CAAAGCGAL-.:

ATGGGCCCTT

CTTCTGTGGT

TTGTGCAGCC

AGGCACCTGT

AGCTGGACAC

CATAAGTGAC TCTTCTGCCC AGCAGTATTA ACTTCACAGA TTCTGCTGAC AAGTTTGAGG

AGTGGAAAGG

TTGCTCTGGG

TGATGTGGAG

CTTGCCAAGG

TGATGACCCT

CATACCATCT

GAGTCCAGCT

GAGCAGGGAG

GGTGGTGTCT

CCACATCACT

TGCTGAGTTT

qGTAGTTAGC 3iC TTTGAA

GAATCC

CACCAACATG

GAAGGAGCTT

AGATGCCTGG

GGTGACCCGA

CTACCTGATA

TCATCCC CTT

AGTCTTC-AGA

GAGCAACAGC

CAAGATCTAG

GAATCTGATC

TCAACCTCTG

GCTGCTCATA

AAGCCAGAAT

GGCCTGACCC

TTAACTAATC

GTGTGTGAAC

TATTTCTGGG

GTCAGAGGAG

CAGGACAGAA

CCCACAGATC

TCATCATTCA

ACAGAGGACA

GAGTGGGTGT

CCCCAGATCC

4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5656 ado CCCACAGCCA CTACTG INFORMATION FOR SEQ ID NO: 2: SEQUENCE CHARACTERISTICS: 50 LENGTH: 5709 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG GTTTCTCAGA TAACTGGGCC CCTGCGCTCA GGAGGCCTTC ACCCTCTGCT CTGGGTAAAG TTCATTGGAA CAGAAAGAAA TGGATTTATC TGCTCTTC' C GTTGAAGAAG TACAAAATGT CATTAATGCT ATGCAGAAAA WO 96/33271 P~U9/52 PCT/US96/05621

TCTTAGAGTG

ATATTTTGCA

CCTTTATGTA

CTTGTTGAA.G

GCAAACAGCT

GTTTCTATCA

CCCGAAAATC

GTGAGAACTC

TTGGGATCTG

CAMAGAATTGT

AAAAAGGCTG

AGTAATAATG

CAGGGTAGTT

TCATTACAGC

GCTGAATTCT

GCTGGAAGTA

CTGAATGCTG

GAGAATCCTA

GTTAATGAGT

GAGTCTGAAT

TATTCTGGTT

TGTAAAAGTG

GGGAAAACCT

ATTATAGGAG

TTAAAGCGTA

GATTTGGCAG

AATGGTCAAG

ATTCAGAATG

ACGAAAGCTG

AATTCAAAAG

GCGCTTGAAC

GATAGTTGTT

TCCCATCTGT

AATTTTGCAT

AGAATGATAT

AGCTATTGAA

ATAATTTTGC

TCCAAAGTAT

CTTCCTTGCA

TGAGGACAAA

ATTCTTCTGA

T'iCAAATCAC

CTTGTGAATT

ATTTGAACAC

CTGTTTCAAA.

ATGAGAACAG

GTAATAAAAG

AGGAAACATG

ATCCCCTGTG

GAGATACTGA

GGTTTTCCAG

CAAATGCCAA

CTTCAGAGAA

AAAGAGTTCA

ATCGGAAGkA

CATTTGTTAC

AXAGGAGACC

TTCAAAAGhC

TGATGAATAT

AGAAAAATCC

AACCTATAAG

CACCTAAAAA

TAGTAGTCAG

CTAGCAGTGA

CTGGAGTTGA

GCTGAAACTT

AACCAAAAGG

AATCATTTGT

AAAAAAGGAA

GGGCTACAGA

GGAAACCAGT

GCAGCGGATA

AGATACCGTT

CCCTCAAGGA

TTCTGAGACG

CACTGAGAAG

CTTGCATGTG

CAGTTTATTA

CAAACAGCCT

TAATGATAGG

TGAGAGAAA

AGATGTTCCT

AAGTGATr(AA AGTAGCTG k'

AATAGACTTA

CTCCAAATCA

GGCAAGCCTC

TGAGCCACAG

TACATCAGGC

TCCTGAAATG

TACTAATAGT

TAACCCAATA

CAGCAGTATA

GAATAGGCTG

TAGAAATCTA

AGAGATAAAG

TCAAGGAACC

CTCAACCAGA

AGCCTACAAG

GCTTTTCAGC

A.ATAACTCTC

AACCGTGCCA

CTCAGTGTCC

CAACCTCAAA

AATAAGGCAA

ACCAGGGATG

GATGTAACAA

CGTGCAGCTG

GAGCCATGTG

CTCACTAAAG

GGCTTAGCAA

CGGACTCCCA

GAATGGAATA

TGGATAACAC,

CT1GrYOPG-.T ,,3A-VTGGACG

CTGGCCAGTG

GTAGAGAGTA

CCCAACTTAA

ATAATACAAG

CTTCATCCTG

ATAAATCAGG

GGTCATGAGA

GAATCACTCG

AGCAATATGG

AGGAGGAAGT

AGCCCACCTA

AAAAAAAAGT

TGTCTCCACA

AGAAAGGGCC

AAAGTACGAG

TTGACACAGG

CTGAACATCT

AAAGACTTCT

AACTCTCTAA

AGACGTCTGT

CTTATTGCAG

AAATCAGTTT

ATACTGAACA

AGAGGCATCC

GCACAAATAC

ACAGAATGAA

GGAGCCAACA

GC. 4 :AGAA.AA

AGCAGAAACT

TAAATAGCAG

i'TGATGACTC

TTCTAAATGA

ATCCTCATGA

ATATU -,',AGA

GCCATGTAAC

AGCGTCCCCT

AGGATTTT.AT

GAACTAACCA

ATAAAACAAA

AAAAAGAATC

AACTCGAATT

CTTCTACCAG

ATTGTACTGA

ACAACCAAAT

GTGTGACCAC

TTCACAGTGT

ATTTAGTCAA

TTTGGAGTAT

AAAAGATGAA

ACAGAGTGAA

CCTTGGAACT

CTACATTGAA

TGTGGGAGAT

GGATTCTGCA

TCATCAACCC

AGAAAAGTAT

TCATGCCAGC

TGTAGAAAAG

TAACAGATGG

AAAGGTAGAT

GCCATGCTCA

CATTCAGAAA

ACATGATGGG

GGTAGATGAA

GGCTTTAATA

CAAAATATTT

TGAAAATCTA

CACAAATAAA

CAAGAAAGCA

AACGGAGCAG

AGGTGATTCT

TGCTTTCAAA

AAATATCCAC

GCATATTCAT

ATTGCAAATT

GCCAGTCAGG

240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 WO 96/33271 WO 9633271PCTIUS96/05621

CACAGCAGAA

AACAAGCCAA

TTAACAAATG

GTCAATCCTA

AATAATGCTG

AGATCTGTAG

AGTATCTCGT

GTGAGTCAGT

AATAGAAATG

GAAACAAGCA

AAGGTTTuU-

TGTGCAACAT

(UkATGTGAAC

ACAGTTAATA

GCCAAATGTA

GAAACTGGAC

CCACTTTTTC

AACTTTGAGG

ACAGTGAGCA

AGCAATATTA

cGrTCCAGTG

GCTATGCTTA

AATTGTAAGC

ACAGATTTCT

GCATCTCAGG

GATACTAGTT

CAGAAAGGAG

TACCGAAGAG

GAGCTTCCCT

AC"'AGGCATA

TCATTGAAGA

GAACATCACC

ACCTACAACT CATGGAAGGT AAAGAT.CCTG CAACTGGAGC CAAGAAGAGT

ATGAACAGAC

CACCTGGTTC

GCCTTCCAAG

AAGACCCCAA

AGAGTAGCAG

TACTGGAAGT

GTGCAGCATT

ACACAGAAGG

TAGAAATr(;A AGCGCCA-7

TCTCTGCCCA

AAAAGGAAGA

TCACTGCAGG

GTATCAAAGG

TCATTACTCC

CCATCAAGTC

AAXCATTCAAT

CAATTAGCCG

ATGAAGTAGG

ATGAAAACAT

GATTAGGGGT

ATCCTGAAAT

CTCCATATCT

TTTGTTCTGA

TTGCTGAAA

AGCTTAGCAG

GGGCCAAGAA

GCTTCCAACA

GCACCGTTGC

ATAGCTTAAA

TTAGTGAGGA

AAGTAAAAGA CATGACAGCG

TTTTACTAAG

AGAAGAAAAA

AGATCTCATG

TATTTCATTG

TAGCACTCTA

TGAMAACCCC

CTTTAAGTAT

AGAAAGTGAA

ATTTGCTCCG

CTCTGGGTCC

AAATCAAGGA

CTTTCCTGTrG

AGGCTCTAGG

AAATAAACAT

ATTTGTTAAA

GTCACCTGAA

TAATAACATT

TTCCAGTACT

TCAAGCAGAA

TTTGCAACCT

AAAAAAGCAA

GATTTCAGAT

GACACCTGAT

TGACATTAAG

GAGTCCTAGC

ATTAGAGTCC

CTTGTTATTC

TACCGAGTGT

TGACTGCAGT

AACAAAATGT

TGT C AAATA

GAAGAGAAAC

TTAAGTGGAG

GTACCTGGTA

GGGAAGGCAA

AAGGGACTAA

CCATTGGGAC

CTTGATGCTC

TTTTCAA.ATC

TTAAAGAAAC

AAGAATGAGT

GTTGGTCAGA

TTTTGTCTAT

GGACTTTTAC

ACTAA.ATGTA

AGAGAAATGG

AGAGAAkAATG

AATGAAGTGG

CTAGGTAGAA

GAGGTCTATA

GAATATGAAG

AACTTAGAAC

GACCTGTTAG

GAAAGTTCTG

CCTTTCACCC

TCAGAAGAGA

GGTAAAGTAA

CTGTCTAAGA

AACCAGGTAA

TCTGCTAGCT

ATACTTTCCC

CCAGTGAACT

TAGAAACAGT

AAAGGGTTTT

CTGATTATGG

AAACAGAACC

TTCATGGTTG

ATGAAGTTAA

AGTATTTGCA

CAGGAAATGC

AAAGTCCAA

CTAATATCAA

&PAGATAAGCC

CATCTCAGTT

AAAACCCATA

AGAAAAATCT

GAAATGAGAA

TTTTTAAAGA

GCTCCAGTAT

ACAGAGGGCC

AACAAAGTCT

AAGTAGTTCA

AGCCTATGGG

ATGATGGTGA

CTGTTTTTAG

ATACACATTT

ACTTATCTAG

ACAATATACC

ACACAGAGGA

TATTGGCAAA

TGTTTTCTTC

AGAGCTGAAG

TAAAGAATTT

TAAAGTGTCT

GCAAACTGAA

CACTCAGGAA

AAATAAATGT

TTCCAA.AGAT

CCACAGTCGG

GAATACATTC

AGAAGAGGAA

AGTCACTTTT

GCCTGTACAG

AGTTGATAAT

CAGAGGCAAC

TCGTATACCA

GCTAGAGGAA

CATTCCAAGT

AGCCAGCTCA

TAATGAAATA

AAAATTGAAT

TCCTGGAAGT

GACTGTTAAT

AAGTAGTCAT

AATAAAGGA.A

CAAA.AGCGTC

GGCTCAGGGT

TGAGGATGAA

TTCTCAGTCT

GAATTTATTA

GGCATCTCAG

ACAGTGCAGT

2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020

A"

WO 96133271 PCTIUS96/05621

GAATTGGAAG

AAACAAATGA

TCAGATGATG

GATTCAAACT

TGCTCAGGGC

CATAACCTGA

GGGAGCCAGC

CTGCGAAATC

GAATACCCTA

GATAGTTCTA

CCATCATTAG

TACCCATCTC

TCTGGGCCAC

CCTTACCTGG

GACAGAGCCC

GTTCCCCAAT

ACTGCTGGGT

TCAACAGAAA

TTTATGCTCG

GAAGAGACTA

AAATATTTTC

TCTATTAAAC

AATGGAAGAA

AGGGGGCTAG

TGGATGGTAC

ACAGGTGTCC

CATGCAATTG

GTAGCACTCT

CACTACTGA

ACTTGACTGC

GGCATCAGTC

AAGAAAGAGG

TAGGTGAAGC

TATCCTCTCA

TAAAGCTCCA

CTTCTAACAG

CAGAACAAAG

TAAGCCAGAA

CCAGTAAAAA

ATGATAGGTG

AAC3AGGAGCT

ACGATTTGAC

AATCTGGAAT

CAGAGTCAGC

TGAAAGTTGC

ATAATGCAAT

GGGTCAACAA

TGTACAAGTT

CTCATGTTGT

TAGGAATTGC

AAAGAAAAAT

ACCACCAAGG

AAATCTOTTG

AGCTGTGTGG

ACCCAATTGT

GGCAGATGTG

AAATACAAAC

TGAAAGCCAG

AACGGGCTTG

AGCATCTGGG

GAGTGACATT

GCAGGAAATG

CTACCCTTCC

CACATCAGAA

TCCAGAAGC

TAA.AGAACCA

GTACATGCAC

CATTAAGGTT

GGAA?.CATCT

CAGCCTCTTC

TCGTGTTGGC

AGAATCTGCC

GGAAGAAAGT

AAGAATGTCC

TGCCAGAAAA

TATGAAAACA

GGGAGGAAAA

GCTGAATGAG

TCCAkAGCGA

CTATGGGCCC

TGCTTCTGTG

GGTTGTGCAG

TGAGGCACCT

Ak-WAGGATC

GGAGTTGGTC

GAAGAAAATA

TGTGAGAGTG

TTAACCACTC

GCTGAACTAG

ATCATAAGTG

AAAGCAGTAT

CTTTCTGCTG

GGAGTGGAAA

AGTTGCTCTG

GTTGATGTGG

TACTTGCCAA

TCTGATGACC

AACATACCAT

CAGAGTCCAG

GTGAGCAGGG

ATGGTGGTGT

CACCACATCA

GATCZTGAGT

TGGGTAGTTA

CATGATTTTG

GCAAGAGAAT

TTCACCAACA

GTGAAGGAGC

CCAGATGCCT

GTGGTGACCC

ACCTACCTGA

CTTTCTTGAT

TGAGTGACAA

ATCAAGAAGA

AAACAAGCGT

AGCAGAGGGA

AAGCTGTGTT

ACTCTTCTGC

TAAC..LTCACA

ACAAGTTTGA

GGTCATCCCC

GGAGTCTTCA

AGGAGCAACA

GGCAAGATCT

CTGAATCTGA

CTTCAACCTC

CTGCTGCTCA

AGAAGCCAGA

CTGGCCTGAC

CTTTAACTAA

TTGTGTGTGA

GCTATTTCTG

AAGTCAGAGG

CCCAGGACAG

TGCCCACAGA

TTTCATCATT

GGACAGAGGA

GAGAGTGGGT

TACCCCAGAT

TGGTTCTTCC

GGAATTGGTT

GCAAAGCATG

CTCTGAAGAC

TACCATGCAA

AGAACAGCAT

CCTTGAGGAC

GAAAAGTAGT

GGTGTCTGCA

TTCTAAATGC

GAATAGAAAC

GCTGGAAGAG

AGAGGGAACC

TCCTTCTGAA

TGCATTGAAA

TACTACTGAT

ATTGACAGCT

CCCAGAAGAA

TCTAATTACT

ACGGACACTG

GGTGACCCAG

AGATGTGGTC

AAGATCTTC

TCAACTGGAA

CACCCTTGGC

CAATGGCTTC

GTTGGACAGT

CCCCCACAGC

4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 510.J 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5709 toLl ACCAGTGCCA GGAGCTGGAC INFORMATION FOR SEQ ID NO:3: Wi SEQUENCE CHARACTERISTICS: LENGTH: 5689 base pairs TYPE: nucleic acid p 21 WO 96/33271 WO 9633271PCTIUS96/05621 STRAflDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: CDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG CCTGCGCTCA GGAGGCCTTC TGGATTTATC TGCTCTTCGC TCTTAGAGTG TCCCATCTGT ACATATTTTG CAAATTTTGC GTCCTTTATG AGCCTACAAG AATCATTTGT GCTTTTCAGC AAAAAAGGAA AATAACTCTC GGGCTACAGA AACCGTGCCA GGAA.ACCAGT CTCAGTGTCC GCAGCGGATA CAACCTCAAA AGATACCGTT AATAAGGCAA CCCTCAAGGA ACCAGGGATG TTCTGAGACG GATGTAACAA CACTGAGAAG CGTGCAGCTG CTTGCA.TGTG GAGCCATGTG CAGTTTATTA CTCACTAAAG CAAACAGCCT GGCTTAGCAA TCAATGATAGG CGGACTCCCA TGAGAGAAAA GAATGGAATA AGATGTTCCT TGGATAACAC AAGTGATGAA CTGTTAGGTT AGTAGCTGAT GTATTGGACG AATAGACTTA CTGGCCAGTG CTCCAAATCA GTAGAGAGTA GGCAAGCCTC CCCAACTTAA TGAGCCACAG ATAATACAAG TACATCAGGC CTTCATCCTG

ACCCTCTGCT

GTTGA.AGAAG

CTGGAGTTGA

ATGCTGAAAC

AAAGTACGAG

TTGACACAGG

CTGAACATCT

AAAGACTTCT

AACTCTC1TAA AGACGTC'2GT

CTTATTGCAG

AAATCAGTTT

ATACTGAACA

AGAGGCATCC

GCACAAATAC

ACAGAATGAA

GGAGCCAACA

GCACAGAAAA

AGCAGAAACT

TAAATAGCAG

CTGATGACTC

TTCTAAATGA

ATCCTCATGA

ATATTGAAGA

GCCATGTAAC

AGCGTCCCCT

AGGATTTTAT

CTGGGT AAAG

TACAAA-ATGT

TCAAGGAACC

TTCTCAACCA

ATTTAGTCAA

TTTGGAGTAT

AAAAGATGAA

ACAGAGTGAA

CCTTGGAACT

CTACATTGAA

TGTGGGAGAT

GGATTCTGCA

TCATCAACCC

AGAAAAGTAT

TCATGCCAGC

TGTAGAAAAG

TAACAGATGG

AAAGGTAGAT

GCCATGCTCA

CATTCAGAAA

ACATGATGGG

GGTAGATGAA

GGCTTTAATA

CAAAATATTT

TGAAAATCTA

CACAAATAAA

CAAGAAAGCA

GTTTCTCAGA

TTCATTGGAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

CTTGTTGAAG

GCAAACAGCT

GTTTCTATCA

CCC-.AAAATC

GTGAGAACTC

TTGGGATCTG

CAAGAATTGT

AAAAAGGCTG

AGTAATAATG

CAGGGTAGTT

TCATTACAGC

GCTGAATTCT

GCTGGAAGTA

CTGAATGCTG

GAGAATCCTA

GTTAATGAGT

GAGTCTGAAT

TATTCTGGTT

TGTAAAAGTG

GGGAAAACCT

ATTATAGGAG

TTAAAGCGTA

GATTTGGCAG

TAACTGGGCC

CAGAAAGAAA

ATGCAGAAAA

A.AGTGTGACC

CCTTCACAGT

AGCTATTGAA

ATAATTTTGC

TCCAAAGTAT

CTTCCTTGCA

TGAGGACAAA

ATTCTTCTGA

TACAAATCAC

CTTGTGAATT

ATTTGAACAC

CTGTTTCAAA

ATGAGAACAG

GTAATAAAAG

AGGAAACATG

ATCCCCTGTG

GAC-ATACTGA

GGTTTTCCAG

CAAATGCCAA

CTTCAGAGAA

AAAGAGTTCA

ATCGGAAGAA

CATTTGTTAC

AAAGGAGACC

TTCAAAAGAC

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 WO 96/3327 1 PCTIUS96/05621

TCCTGAAATG

TACTAATAGT

TAACCCAATA

CAGCAGTATA

GAATAGGCTG

TAGAAATCTA

AGAGATAAAG

CATGGAAGGT

AAGTAAAAGA

TTTTACTAAG

AGAAGAAAAA

AGATCTCATG

TATTTCATTG

TAGCACTCTA

TGAAAACCCC

CTTTAAGTAT

AGAAAGTGAA

ATTTGCTCCG

CTCTGGGTCC

AAATCAAGGA

CTTTCCTGTG

AGGCTCTAGG

AAATAAACAT

ATTTGTTAAA

GTCACCTGAA

TAATAACATT

TTCCAGTACT

TCAAGCAGAA

TTTGCAACCT

AAAAAAGCAA

GATTTCAGAT

GACACCTGAT

ATAAATCAGG

GGTCATGAGA

GAATCACTCG

AGCAATATGG

AGGAGGAAGT

AGCCCACCTA

AAAAAAAGT

AAAGAACCTG

CATGACAGCG

TGTTCAAATA

GAAGAGAAAC

TTAAGTGGAG

GTACCTGGTA

GGGAAGGCA

AAGGGACTAA

CCATTGGGAC

CTTGATGCTC

TTTTCAAATC

TTAAAGAAAC

AAGAATGAGT

GTTGGTCAGA

TTTTGTCTAT

GGACTTTTAC

ACTAAATGTA

AGAGAAATGG

AGAGAAAATG

AATGAAGTGG

CTAGGTAGAA

GAGGTCTATA

GAATATGAAG

AACTTAGAAC

GACCTGTTAG

GAACTAACCA

ATAAA.AC:AA

AAAAAGAATC

AACTCGAATT

CTTCTACCAG

ATTGTACTGA

ACAACCAAAT

CAACTGGAGC

ATACTTTCCC

CCAGTGAACT

TAGAAACAGT

AAAGGGTTTT

CTGATTATGG

AAACAGAACC

TTCATGGTTG

ATGAAGTTAA

AGTATTTGCA

CAGGAAATGC

AAAGTCCAAA

CTAATATCAA

AAGATAAGCC

CATCTCAGTT

AAAACCCATA

AGAAAAATCT

GAAATGAGA-k

TTTTTAAAGA

GCTCCAGTAT

ACAGAGGGCC

AACAAAGTCT

AAGTAGTTCA

AGCCTATGGG

ATGATGGTGA

AACGGAGCAG AATGGTCAAG AGGTGATTCT ATTCAGAATG TGCTTTCAAA ACGAAAGCTG AAATATCCAC PATTCAAAAG

GCATATTCAT

ATTGCAJAATT

GCCAGTCAGG

CAAGAAGAGT

AGAGCTGAAG

TAAAGAATTT

TAAAGTGTCT

GCAAACTGAA

CACTCAGGAA

AAATAAATGT

TTCCAAAGAT

CCACAGTCGG

GAATACATTC

AGAAGAGGAA

AGTCACTTTT

GCCTGTACAG

AGTTGATAAT

CAGAGGCAAC

TCGTATACCA

GCTAGAGGAA

CATTCCAAGT

AGCCAGCTCA

TAATGAAATA

AAAATTGAAT

TCCTGGAAGT

GACTGTTAAT

AAGTAGTCAT

AATAAAGGAA

GCGCTTGAAC

GATAGTTGTT

CACAGCAGAA

AACAAGCCAA

TTAACAAATG

GTCAATCCTA

AATAATGCTG

AGATCTGTAG

AGTATCTCGT

GTGAGTCAGT

AATAGAAATG

GAAACAAGCA

AAGGTTTCAA

TGTGCAACAT

GAATGTGAAC

ACAGTTAATA

GCCAAATGTA

GAAACTGGAC

CCACTT'TTC

AACTTTGAGG

ACAGTGAGCA

AGCAATATTA

GGTTCCAGTG

GCTATGCTTA

AATTGTAAGC

ACAGATTTCT

GCATCTCAGG

GATACTAGTT

TGATGAATAT

AGAAAAATCC

AACCTATAAG

CACCTAAAAA

TAGTAGTCAG

CTAGCAGTGA

ACCTACAACT

ATGAACAGAC

CACCTGGTTC

GCCTTCCAAG

A.AGACCCCAA

AGAGTAGCAG

TACTGGAAGT

GTGCAGCATT

ACACAGAAGG

TAGAAATGGA

AGCGCCAGTC

TCTCTGCCCA

AAAAGGAAGA

TCACTGCAGG

GTATCAAAGG

TCATTACTCC

CCATCAAGTC

AACATTCAAT

CAATTAGCCG

ATGAAGTAGG

ATGAAAkACAT

GATTAGGGGT

ATCCTGAAAT

CTCCATATCT

TTTGTTCTGA

TTGCTGAAAA

1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 WO 96/33271 PTU9/52 PCTIUS96105621

TGACATTAAG

GAGTCCTAGC

ATTAGAGTCC

CTTGTTATTT

TACCGAGTGT

TGACTGCAGT

AACAAAATGT

AAATACAAAC

TGAAAGCCAG

AACGGGCTTG

AGCATCTGGG

GAGTGACATT

GCAGGAAATG

CTACCCTTCC

CACATCZLGAA

TCCAGAAGGC

TAAAACCA

GTACATGCAC

CATTAAGGTT

GGAAACATCT

CAGCCTCTTC

TCGTGTTGGC

AGAATfCTGCC

GGAAGAAAGT

A.AGAATGTCC

TGCCAGAAAA

TATGAAAACA

GGGAGGAAAA

GCTGAATGAG

TCCAAAGCGA

CTATGGGCCC

TGCTTCTGTG

GAAAGTTCTG

CCTTTCACCC

TCAGAAGAGA

GGTA.AAGTAA

C .'GTCTAAGA

AACCAGGTAA

TCTGCTAGCT

ACCCAGGATC

GGAGTTGGTC

GAAGAAAATA

TGTGAGAGTG

TTAACCACTC

GCTGAACTAG

ATCATAAGTG

AAAGCAGTAT

CTTTCTGCTG

GGAGTCGAA

AGTTGCTCTG

GTTGATGTGG

TACTTGCCAA

TCTGATGACC

AACATACCAT

CAGAGTCCAG

GTGAGCAGGG

ATGGTGGTGT

CACCACATCA

GATGCTGAGT

TGGGTAGTTA

CATGATTTTG

GCAAGAGAAT

TTCACCAACA

GT1GAAGGAGC

CTGTTTTTAG

ATACACATTT

ACTTATCTAG

ACAATATACC

ACACAGAGGA

TATTGGCAAA

TGTTTTCTTC

CTTTCTTGAT

TGAGTGACAA

ATCAAGAAGA

AAACAAGCGT

AGCAGAGGGA

A.AGCTGTGTT

ACTCTTCTGC

TAACTTCACA

ACAAGTTTGA

GGTCATCCCC

GGAGTCTTCA

AGGAGCAACA

GGCAAGATCT

CTGAATCTGA

CTTCAACCTC

CTGCTGCTCA

AGAAGCCAGA

CTGGCCTGAC

CTTTAACTAA

TTGTGTGTGA

GCTATTTCTG

AAGTCAGAGG

CCCAGGACAG

TGCCCACAGA

TTTCATCATT

CAAAAGCGTC

GGCTCAGGGT

TGAGGATGAA

TTCTCAGTCT

GAATTTATTA

GGCATCTCAG

ACAGTGCAGT

TGGTTCTTCC

GGAATTGGTT

GCAAAGCATG

CTCTGAAGAC

TACCATGCAA

AGAACAGCAT

CCTTGAGGAC

GAAAAGTAGT

GGTGTCTGCA

TTCTAAATGC

GAATAGAAAC

GCTGGAAGAG

AGAGGGAACC

TCCTTCTGAA

TGCATTG.AA

TACTArTGAT

ATTGACAGCT

CCCAGAAGAA

TCTAATTACT

ACGGACACTG

GGTGACCCAG

AGATGTGGTC

AAAGATCTTC

TCAACTGGAA

CACCCTTGGC

CAGAA.AGGAG

TACCGAAGAG

GAGCTTCCCT

ACTAGGCATA

TCATTGAAGA

GAACATCACC

Gi-, ,TGGAAG

AACAAATGA

TCAGATGATG

GATTCAAACT

TGCTCAGGGC

CATAACCTGA

GGGAGCCAGC

CTGCGAAATC

GAAIACCCTA

GATAGTTCTA

CCATCATTAG

TACCCATCTC

TCTGGGCCAC

CCTTACCTGG

GACAGAGCCC

GTTCCCCAAT

ACTGCTGGGT

TCAACAGAAA

TTTATGCTCG

GAAGAGACTA

AAATATTTTC

TCTATTAAAG

AATGGAAGAA

AGGGGGCTAG

TGGATGGTAC

ACAGGTGTCC

AGCTTAGCAG

GGGCCAAGAA

GCTTCCAACA

GCACCGTTGC

ATAGCTTAAA

TTAGTGAGGA

ACTTGACTGC

GGCATCAGTC

AAGAAAGAGG

TAGGTGAAGC

TATCCTCTCA

TAAAGCTCCA

CTTCTAACAG

CAGAACAAAG

TAAGCCAGAA

CCAGTAAAAA

ATGATAGGTG

AAGAGGAGCT

ACGATTTGAC

AATCTGGAAT

CAGAGTCAGC

TGAAAGTTGC

ATAATGCAAT

GGGTCAACAA

TGTACAAGTT

CTCATGTTGT

TAGGAATTGC

AAAGAAAAAT

AC CACCAAGG

AAATCTGTTG

.AGCTGIGTGG

ACCCAATTGT

3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 Ii' p WO 96/33271

PCTJUS

GGTTGTGCAG CCAGATGCCT GGACAGAGGA CA.ATGGCTTC CATGCAATTG GGCAGATGTG TGAGGCACCT GTGGTGACCC GAGAGTGGGT GTTGGACAGT GTAGCACTCT ACCAGTGCCA GGAGCTGGAC ACCTP.CCTGA TACCCCAGAT CCCCCACAGC CACTACTGA INFORMATION F I SEQ ID NO: 4: SEQUENCE CHARACTERISTICS: LENGTH: 5711 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: 96/05621 AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG

CCTGCGCTCA

TGGATTTATC

TCTTAGAGTG

ACATATTTTG

GTCCTC2TATG

AACTTGTTGA.

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AATTGGGATC

?-CAAGAATT

CAAAAAAGGC

CCAGTAATAA

ATCAGGGTAG

GCTCATTACA

AGGCTGAATT

GGGCTGGAAG

ATCTGAIXTGC

CAGAGAATCC

AAGTTAATGA

GGGAGTCTGA

GGAGGCCTTC

TGCTCTTCGC

TCCCATCTGT

CAAATTTTGC

TAAGAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAGGACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGA'

TGATTTGAAC

TTCTGTTTCA

GCATGAGAAC

CTGTAATAAA

TAAGGAAACA

TGATCCCCTG

TAGAGATACT

GTGGTTTTCC

ATCAAATGCC

ACCCTCTGCT

GTTGAAGAAG

CTGGAGTTGA

ATGCTGAAAC

ATAACCAAAA

AAAATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

AAGCAGCGGA

GAAGATACCG

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

TGTAATGAT.A

TGTGAGAGAA

GAAGATGTTC

Af"7AAGTGATG

AAAGTAGCTG

CTGGGTAAAG

T.ACAAAATGT

TCAAGGAACC

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAI)ACCGTGC

GTCTCAGTGT

TACAACCTCA

TTAATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAGC

GGCGGACTCC

AAGAATGGAA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

GTTTCTCAGA

TTCATC-rAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGAACAT

CAAAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAA.AT

AGACAGAATG

AAGGAGCCAA

CAGCACAGAA

TAAGCAGAAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAA.AT

TAAC'rGGGCC

CAGAAAGAAA

ATGCAGAAAA

AAGTGTGACC

ccir~CACAGG

AGATTTAGTC

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

A.NCCTTGGAA

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

AAAAAGGTAG

CTGCCATGCT

AGCATTCAGA

TCACATGATG

GAGGTAGATG

5580 5640 5689 120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1330

I

Al- 4) WO 96/33271 PCTIUS96/05621

AATATTCTGG

TATGTAAAAG

TTGGGAAAAC

TAATTATAGG

AATTAAAGCG

CAGATTTGGC

AGAATGGTCA

CTATTCAGAA

AAACGAAAGC

ACAATTCAAA

ATGCGCTTGA

TTGATAGTTG

GGCACAGCAG

GTAACAAGCC

AGTTAACAAA

TTGTCAATCC

CTAATAATGr AAAGATCTGr

AAAGTATCTC

GTGTGAGTCA

ATAATAGAAA

GGGAAACAAG

TCAAGGTTTC

AATGTGCAAC

TTGAATGTGA

AGACAGTTAA

ATGCCAAATG

ACGAAACTGG

CACCACTTTT

AAAACTTTGA

GTACAGTGAG

CAAGCAATAT

TTCTTCAGAG

TGA.AAGAGTT

CTATCGGAAG

AGCATTTGTT

TAAAAGGAGA

AGTTCAAAAG

AGTGATG.AAT

TGAGAAAAAT

TGAACCTATA

AGCACCTAAA

ACTAGTAGTC

TTCTAGCAGT

AAACCTACAA

AAATGJAACAG

TGCACCTGGT

TAGCCTTCCA

TGAAGACCCC

AGAGAGTAGC

GTTACTGGAA

GTGTGCAGCA

TGACACAGAA

CATAGAAATG

AAAGCGCCAG

ATTCTCTGCC

ACAAAAGGAA

TATCACTGCA

TAGTATCAAA

ACTCATTACT

TCCCATCAAG

GGAACATTCA

CACAATTAGC

TAATGAAGTA

AAAATAGACT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCAG

ACTCCTGAA.

ATTACTAATA

CCTAACCCAA

AGCAGCAGTA

AAGAATAGGC

AGTAGAAATC

GAAGAGATAA

CTCATGGAAG

ACAAGTAAAA

TCTTTTACTA

AGAGAAGAAA

AAAGATCTCA

AGTATTTCAT

GTTAGCACTC

TTTGAAAACC

GGCTTTAAGT

GAAGAAAGTG

TCATTTGCTC

CACTCTGGGT

GAAAATCA.AG

GGCTTTCCTG

GGAGGCTCTA

CCAAATAAAC

TCATTTGTTA

ATGTCACCTG

CGTAATAACA

GGTTCCAGTA

TACTGGCCAG

CAGTAGAGAG

TCCCCAACTT

AGATAATACA

GCCTTCATCC

TGATAAATCA

GTGGTCATGA

TAGAATCACT

TAAGCAATAT

TGAGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GTAAAGAACC

GACATGACAG

AGTGTTCAAA

AAGAAGAGA

TGTTAAGTGG

TGGTACCTC'

TAGGGAAGGC

CCAAGGGACT

ATCCATTGGG

AACTTGATGC

CGTTTTCAAA

CCTTAAAGAA

GAAAGAATGA

TGGTTGGTCA

GGTPTTTGTCT

ATGGACTTTT

AAACTAAATG

APIAGAGAAP.T

TTAGAGkAAA

CTAMOOAIT

TGATCCTCAT

TAATATTGAA

AAGCCATGTA

AGAGCGTCCC

TGAGGATTTT

GGGAACTAAC

GAATAAAACA

CGAAAA.AGAA

GGAACTCGAA

GTCTTCTACC

TAATTGTACT

GTACAACCAA

TGCAACTGGA

CGATACTYTC

TACCAGTGAA

ACTAGAAACA

AGAAAGGGTT

TACTGATTAT

AAAAACAGAA

AATTCATGGT

ACATGAAGTT

TCAGTATTTG

TCCAGGAALAT

ACAAAGTCCA

GTCTAATA'!L

GAAAGATAAG

ATCATCTCAG

ACAAAACCCA

TAAGAAAAAT

GGGAAATGAG

TGTTTTTAAA

GGGCTCCAGT

GAGGCTTTAA

GACAAAATAT

ACTGAAAATC

CTCACAAATA

ATCAAGAAAG

CAAACGGAGC

AAAGGTGATT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GAATTGCAAA

ATGCCAGTCA

GCCAAGAAGA

CCAGAGCTuA

CTTAAAGAAT

GTTAAAGTGT

TTGCAAACTG

GGCACTCAGG

CCAAATAAAT

TGTTCCAAAG

AACCACAGTC

CAGAATACAT

GCAGAAGAGG

AAAGTCACTT

AAGCCTGTAC

CCAGTTGATA

TTCAGAGGCA

TATCGTATAC

CTGCTAGAGG

AACATTCCAA

GAAGCCAGCT

ATTAATGAAA

1440 1500 1560 1620 :680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300

U

13 WO 96/33271 PCTIUS96/05621

TAGGTTCCAG

ATGCTATGCT

GTAATTGTAA

ATACAGATTT

ATGCATCTCA

AAGATACTAG

TCCAGAAAGG

GTTACCGAAG

AAGAGCTTCC

CTACTAGGCA

TATCATTGAA

AGGAACATCA

GTGAATTGGA

CCAAACAAAT

TTTCAGATGA

TGGATTCAAA

ACTGCTCAGG

AACATAACCT

ATGGGAGCCA

ACCTGCGAAA

GTGAATACCC

CAGATAGTTC

GCCCATCATT

ACTACCCATC

AGTCTGGGCC

CCCCTTACCT

AAGACAGAGC

AAGTTCCCCA

ATACTGCTGG

CTTCAACAGA

AATTTATGCT

CTGAAGAGAC

TGATGAAAAC

TAGATTAGGG

GCA','CCTGAA

CTCTCCATAT

GGTTTGTTCT

TTTTGCTGAA

AGAGCTTAGC

AGGGGCCAAG

CTGCTTCCAA

TAG-CACCGTT

GAATAGCTTA

CCTTAGTGAG

AGACTTGACT

GAGGCATCAG

TGAAGAAAGA

CTTAGGTGAA

GCTATCCTCT

GATAAAGCTC

GCCTTCTAAC

TCCAGAACAA

TATAAGCCAG

TACCAGTAAA

AGATGATAGG

TCAAGAGGAG

A::ACGATTTG

GGAATCTGGA

CCCAGAGTCA

ATTGAAAGTT

GTATAATGCA

AAGGGTCAAC

CGTGTACAAG

TACTCATGTT

ATTCAAGCAG

GTTTTGCAAC

ATAAAAAGC

CTGATTTCAG

GAGACACCTG

A-ATGACATTA

AGGAGTCCTA

AAAJTAGAGT

CACTTGTTAT

GCTACCGAGT

AATGACTGCA

GAAACAAAiAT

GCAAATA'CAA

TCTGAMVAGCC

GGAACGGGCT

GCAGCATCTG

CAGAGTGACA

CAGCAGGAAA

AGCTACCCTT

AGU CATCAG

AATCCAGAAG

AATAAAGAAC

TGGTACATGC

CTCATTAAGG

ACGGAAACAT

ATCAGCCTCT

GCTCGTGTTG

GCAGAATCTG

ATGGAA jAAA

AAAAGP.ATGT

AACTAGGTAG

CTGAGGTCTA

AAGAATATGA

ATAACTTAGA

ATGACCTGTT

AGGAAAGTTC

GCCCTTTCAC

CCTCAGA.AGA

TTGGTAAAGT

GTCTGTCTAA

GTAACCAGGT

GTTCTGCTAG

ACACCCAGGA

AGGGAGTTGG

TGGAAGAAA.A

GGTGTGAGAG

TTTTAACCAC

TGGCTGAACT

CCATCATAAG

AAAAAGCAGT

GCC'rTTCTGC

CAGGAGTGGA

ACAGTTGCTC

TTGTTGATGT

CTTACTTGCC

TCTCTGATGA

GCAACATACC

CCCAGAGTCC

GTGTGAGCAG

CCATGGTGGT

AAACAGAGGG

TAAACAAAGT

AGAAGTAGTT

ACAGCCTATG

AGATGATGGT

TCCOTGTTTTT

CCATACACAT

GAACTTATCT

AAACAATATA

GAACACAGAG

AATATTGGCA

CTTGTTTTCT

TCC TTTCTTG

TCTGAGTGAC

TAATCAAGAA

TGAAACAAGC

TCAGCAGAGG

AGAAGCTGTG

TGACTCTTCT

ATTAACTTCA

TGACAAGTTT

AAGGTCATCC

TGGGAGTCTT

GGAGGAGCAA

AAGGCAAGAT

CCCTGAATCT

ATCTTCAACC

AGCTGCTGCT

GGAGAAGCCA

GTCTGGCCTG

CACTTTAACT

GTTTGTGTGT

CCAAAATTGA

CTTCCTGGAA

CAGACTGTTA

GGAAGTAGTC

GAAATAAAGG

AGCAAAAGCG

TTGGCTCAGG

AGTGAGGATG

CCTTC'TCAGT

GAGAATTTAT

AAGGCATCTC

TCACAGTGCA

ATTGGTTCTT

AAGGAATTGG

GAGCAAAGCA

GTCTCTGAAG

GATACCATGC

TTAGAACAGC

GCCCTTGAGG

CAGAAAAGTA

GAGGTGTCTG

CCTrTCTAAAT

CAGAATAGAA

CAGCTGGAAG

CTAGAGGGAA

GATCCTTCTG

TCTGCATTGA

CATACTACTG

GAATTGACAG

ACCCCAGAAG

AATCTAATTA

GAACGGACAC

3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4 2 0 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 TTTGCCAGAA AACACCACAT GTTATG-AAAA CAGATGCTGA 27 WO 9633271PCT[US96/05621 WO 96/33271 TGAAATATTT TCTAGGAATT GCGGGAGGAA 'LiU(G±AGT± TTAX ±G ICAC AGTCTATTAA AGAA.AGAA.AA ATGCTGAATG AGCATGATTT TGAAGTCAGA GGAGATGTGC TCAATGGAAG AAACCACCAA GGTCCAAAGC GAGCAAGAGA ATCCCAGGAC AGAAAGATC TCAGGGGGCT AGAAATCTGT TGCTATGGGC CCTTCACCAA CATGCCCACA GATCAACTGC AATGGATGGT ACAGCTGTGT GGTGCTTCTG TGGTGAAGGA GCTTTCATCA TTCACCCTT( GCACAGGTGT CCACCCAATT GTGGTTGTGC AGCCAGATGC CTGGACAGAG GACAATGGC: TCCATGCAAT TGGGCAGATC TGTGAGGCAC CTGTGGTGAC CCGAGAGTGG GTGTTGGAC~ GTGTAGCACT CTACCAGTGC CAGGAGCTGG ACACCTACCT GATACCCCAG ATCCCCCAC2 GCCACTACTG A INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 59 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID TGTCCTTAAA AGGTTGATAA TCACTTGCTG AGTGTGTTTC TCAAACAAGT TAATTTCAG INFORMATION FOR SEQ ID NO:6: SEQUENCE CHARACTERISTICS: LENGTH: 5710 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ 11) NO:6:

I,

5280 5340 5400 5460 5520 5580 5640 5700 5711 59 AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG

CCTGCGCTCA

TGGATTTATC

TCTTAGAGTG

ACATATTTTG

GTCCTTTATG

AACTTGTTGA

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

GGAGGCCTTC

TGCTCTTCGC

TCCCATCTGT

CAAATTTTGC

TAAGAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

ACCCTCTGCT

GTTGAAGAAG

CTGGAGTTGA

ATGCTGAAAC

ATAACCAAAA

AAAATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

CTGGGTAAAG

TACAAAATGT

TCAAGGAACC

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAACCGTGC

GTCTCAGTGT

GTTTCTCAGA

TTCAT~nGAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGAACAT

CAAAAGACTT

CCAACTCTCT

TAACTGGGCC

CAGAAAGAAA

ATGCAGAAAA

A.AGTGTGACC

CCTTCACAGT

AGATTTAGTC

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

AACCTTGGAA

I alp, 4011 zt7t WO 96/3327 1 PCTJUS96/05621

CTGTGAGAAC

AATTGGGATC

ATCAAGAATT

CAAAAAAGGC

CCAGTAATAA

A'CAGGGTAG

GCTCATTACA

AGGCTGAATT

GGCTGGAAGT

TCTGAATGCT

kGAGAATCCT rGTTAATGAG

GGAGTCTGAA

ATATTCTGGT

ATGTAAA.AGT

TGGGAAAACC

AATTATAGGA

ATTAAAGCGT

AGATTTGGCA

GAATGGTCAA

TATTCAGAAT

AACGAAAGCT

CAATTCAAAA

TGCGCTTGAA

TGATAGTTGT

GCACAGCAGA

TAACAAGCCA

GTTAACAAAT

TGTCAATCCT

TAATAATGCT

AAGATCTGTA

TCTGAGGACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGAA

TGATTTGAAC

TTCTGTTTCA

GCATGAGAAC

CTGTAATAA.A

AAGGAAACAT

GATCCCCTGT

AGAGATACTG

TGGTTTTCCA

TCAAATGCCA

TCTTCAGAGA

GAAAGAGTTC

TATCGGAAGA

GCATTTGTTA

AAAAGGAGAC

GTTCAAAAGA

GTGATGAATA

GAGAAAAATC

GAACCTATAA

GCACCTAAAA

CTAGTAGTCA

TCTAGCAGTG

AACCTACAAC

AATGAACAGA

GCACCTGGTT

AGCCTTCCAA

GAAGACCCCA

GAGAGTAGCA

AAGCfLGCGGA

GAAGATACCG

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACGCC

GTAATGATAG

GTGAGAGAAA

AAGATGTTCC

GAAGTGATGA

AAGTAGCTGA

AAATAGACTT

ACTCCAAATC

AGGCAAGCCT

CTGAGCCACA

CTACATCAGG

CTCCTGAAAT

TTACTAATAG

CTAACCCAAT

GCAGCAGTAT

AGAATAGGCT

GTAGAAATCT

AAGAGATAAA

TCATGGAAGG

CAAGTAAAAG

CTTTTACTAA

GAGA.AGAAAA

AAGATCTCAT

GTATTTCATT

TACAACCTCA

TTAATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

TGGCTTAGCA

GCGGACTCCC

AGAATGGAAT

TTGGATAACA

ACTGTTAGGT

TGTATTGGAC

ACTGGCCAGT

AGTAGAGAGT

CCCCAACTTA

GATAATACA6A

CCTTCATCCT

GATAAATCAG

TGGTCATGAG

AGAATCACTC

AAGCALATATG

GAGGAGGAAG

AAGCCCACCT

GAAAAAAAAG

TAAAGAACCT

ACATGACAGC

GTGTTCAAAT

AGAAGAGAAA

GTTAAGTGGA

GGTACCTGGT

AAAGACGTCT

AACTTATTGC

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAALAT

AGACAGAATG

AGGAGCCAAC

AGCACAGAAA

AAGCAGAAAC

CTAAATAGCA

TCTGATGACT

GTTCTAAATG

GATCCTCATG

A6ATATTGAAG

AGCCATGTAA

GAGCGTCCCC

GAGGATTTTA

GGAACTAAC'7

AATAAAACAA

GAAAAAGAAT

GAAC-TCGAAT

TCTTCTACCA

AATTGTACTG

TACAACCAAA

GCAACTGGAG

GATACTTTCC

ACCAGTGAAC

CTAGAAACAG

GAAAGGGTTT

ACTGATTATG

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

ATAACAGATG

AAAAGGTAGA

TGCCATGCTC

GCATTCAGAA

CACATGATGG

AGGTAGATGA

AGGCTTTAAT

ACAAAATATT

CTGAAAATCT

TCACAA6ATAA

TCAAGAA.AGC

AA6ACGGAGCA zAGGTGATTC

CTGCTTTCAA

TAALATATCCA

GGCATATTCA

AATTGCAA.AT

TGCCAGTCAG

CCAAGAAGAG

CAGAGCTGA.A

TTAAAGAATT

TTAAAGTGTC

TGCAAACTGA

GCACTCAGGA

660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 AAGTATCTCG TTACTGGAAG TTAGCACTCT AGGGAAGGCA AAAACAGAAC CAAATAAATG WO 96/3327 1 PCTIUS96/05621

TGTGAGTCAG

TAATAGAAAT

GGAAACAAGC

CAAGGTTTCA

ATGTGCAACA

TGAATGTGAA

GACAGTTAAT

TGCCAAATGT

CGAAACTGGA

ACCACTTTTT

AAACTTTGAG

TACAGTGAGC

AAGCAATATT

AGGTTCCAGT

TGCTATGCTT

TAATTGTAAG

TACAGATTTC

TGCATCTCAG

AGATACTAGT

CCAGAAAGGA

TTACCGAAGA

AGAGCTTCCC

TACTAGGCAT

ATCATTGAAG

GGAACATCAC

TGAATTGGAA

CAAACAAATG

TTCAGATGAT

GGATTCAAAC

CTGCTCAGGG

ACATAACCTG

TGTGCAGCAT

GACACAGAAG

ATAGAAATGG

A.AGCGCCAGT

TTCTCTGCCC

CAAAAGGAAG

ATCACTGCAG

AGTATCAAAG

CTCATTACTC

CCCATCAAGT

GAACATTCAA

ACAATTAGCC

AATGAAGTAG

GATGAAAACA

AGATTAGGGG

CATCCTGAAA

TCTCCATATC

GTTTGTTCTG

TTTGCTGAAA

GAGCTTkGCA

GGGGCCAAGA

TGCTTCCAAC

AGCACCGTTG

AATAGCTTAA

CTTAGTGAGG

GACT'TGACTG

AGGCATCAGT

GAAGAAAGAG

TTAGGTGAAG

CTATCCTCTC

ATAAAGCTCC

TTGAAAACCC

GCTTTAAGTA

AAGA7AAGTGA

CATTTGCTCC

ACTCTGGGTC

AAAATCAAGG

GCTTTCCTGT

GAGGCTCTAG

CAAATAAACA

CATTTGTTAA

TGTCACCTGA

GTAATAACAT

GTTCCAGTAC

TTCAAGCAGA

TTTTGCAACC

TAAAAAAGCA

TGATTTCAGA

AGACACCTGA

ATGACATTAA

GGAGTCCTAG

AATTAGAGTC

ACTTGTTATT

CTACCGAGTG

ATGACTGCAG

AAACAAAATG

CAAATACAAA

CTGAAAGCCA

GAACGG'-'TT

CAGCATCTGG

AGAGTGACAT

AGCAGGAAAT

CAAGGGACTA

TCCATTGGGA

ACTTGATGCT

GTTTTCAAAT

CTTAAAGAAA

AAAGAATGAG

GGTTGGTCAG

GTTTTGTCTA

TGGACTTTTA

AACTAAATGT

AAGAGAAATG

TAGAGAAAAT

TA.ATGAAGTG

ATTCATGGTT

CATGAAGTTA

CAGTATTTGC

CCAGGAAATG

CAAAGTCCAA

TCTAATATCA

AAAGATAAGC

TCATCTCAGT

CAAAACCCAT

AAGAAAAATC

GGAAATGAGA

GTTTTTAAAG

GGCTCCAGTA

ACTAGGTAGA AACAGAGGGC TGAGGTCTAT AAACAAAGTC AGAATATGAA GAAGTAGTTC

GTTCCAAAGA

ACCACAGTCG

AGAATACATT

CAGAAGAGGA

AAGTCACTTT

AGCCTGTACA

CAGTTGATAA

TCAGAGGCAA

ATCGTATACC

TGCTAGAGGA

ACATTCCAAG

AAGCCAGCTC

TTAATGAAAT

CAAAATTGA.A

TTCCTGGAAG

AGACTGTTAA

GAAGTAGTCA

AAATAAAGGA

GCAAAAGCGT

TGGCTCAGGG

GTGAGGATGA

CTTCTCAGTC

AGAATTTATT

AGGCATCTCA

CACAGTGCAG

TTGGTTCTTC

AGGAATTGGT

AGCAA.AGCAT

TCTCTGAAGA

ATACCATGCA

TAGAACAGCA

TAACTTAGAA

TGACCTGTTA

GGAAAGTTCT

CCCTTTCACC

CTCAGAAGAG

TGGTAAAGTA

TCTGTCTAAG

TAACCAGGTA

TTCTGCTAGC

CACCCAGGAT

GGGAGTTGGT

GGAAGAAAAT

GTGTGAGAGT

TTTAACCACT

GGCTGAACTA

CAGCCTATGG

GATGATGGTG

GCTGTTTTTA

CATACACATT

AACTTATCTA

AACAATATAC

AACACAGAGG

ATATTGGCAA

TTGTTTTCTT

CCTTTCTTGA

CTGAGTGACA

AATCAAGAAG

GAAACAAGCG

CAGCAGAGGG

GAAGCTGTGT

2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 TGGGAGCCAG CCTTCTAACA GCTACCCTTC CATCATAAGT GACTCTTCTG CCCTTGAGGA

'I

WO 96/33271 PCT/US96/05621

CCTGCGAAAT

TGAATACCCT

AGATAGTTCT

CCCATCATTA

CTACCCATCT

GTCTGGGCCA

CCCTTACCTG

AGACAGAGCC

AGTTCCCCAA

TACTGCTGGG

TTCAACAGAA

ATTTATGCTC

TGAAGAGACT

GAAATATTTT

GTCTATTAAA

CAATGGAAGA

CAGGGGGCTA

ATGGATGGTA

CACAGGTGTC

CCATGCAATT

TGTAGCACTC

CCAGAACAAA

ATAAGCCAGA

ACCAGTAAAA

GATGATAGGT

CAAGAGGAGC

CACGATTTGA

GAATCTGGAA

CCAGAGTCAG

TTGAAAGTTG

TATAATGCAA

AGGGTCAACA

GTGTACAAGT

ACTCATGTTG

CTAGGAATTG

GAAAGAAAAA

AACCACCAAG

GAAATCTGTT

CAGCTGTGTG

CACCCAATTG

GGGCAGATGT

TACCAGTGCC

GCACATCAGA

ATCCAGAAGG

ATAAAGAACC

GGTACATGCA

TCATTAAGGT

CGGAAACATC

TCAGCCTCTT

CTCGTGTTGG

CAGAATCTGC

TGGAAGAAAG

AAAGAATGTC

TTGCCAGAAA

TTATGAAAAC

CGGGAGGAAA

TGCTGAATGA

GTCCAAAGCG

GCTATGGGCC

GTGCTTCTGT

TGGTTGTGCA

GTGAGGCACC

AGGAGCTGGA

AAAAGCAGTA

CCTTTCTGCT

AGGAGTGGAA

CAGTTGCTCT

TGTTGATGTG

TTACTTGCCA

CTCTGATGAC

CAACATACCA

CCAGAGTCCA

TGTGAGCAGG

CATGGTGGTG

ACACCACATC

AGATGCTGAG

ATGGGTAGTT

GCATGATTTT

AGCAAGAGAA

CTTCACCAAC

GGTGAAGGAG

GCCAGATGCC

TGTGGTGACC

CACCTACCTG

TTAACTTCAC

GACAAGTTTG

AGGTCATCCC

GGGAGTCTTC

GAGGAGCAAC

AGGCAAGATC

CCTGAATCTG

TCTTCAACCT

GCTGCTGCTC

GAGAAGCCAG

TCTGGCCTGA

ACTTTAACTA

TTTGTGTGTG

AGCTATTTCT

GAAGTCAGAG

TCCCAGGACA

ATGCCCACAG

CTTTCATCAT

TGGACAGAGG

CGAGAGTGGG

ATACCCCAGA

AGAAAAGTAG

AGGTGTCTGC

CTTCTAAATG

AGAATAGAAA

AGCTGGAAGA

TAGAGGGAAC

ATCCTTCTGA

CTGCATTGAA

ATACTACTGA

AATTGACAGC

CCCCAGAAGA

ATCTAATTAC

AACGGACACT

GGGTGACCCA

GAGATGTGGT

GAAAGATCTT

ATCAACTGGA

TCACCCTTGG

ACAATGGCTT

TGTTGGACAG

TCCCCCACAG

4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5710

U

J7

CCACTACTGA

INFORMATION FOR SEQ ID NO:7: SEQUENCE CHARACTERISTICS: LENGTH: 5709 base pairs TYPE: nucleic acid 50 STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA 55 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG GTTTCTCAGA. TAACTGGGCC CCTGCGCTCA GGAGGCCTTC ACCCTCTGCT CTGGGTAAAG TTCATTGGAA CAGAAAGAAA TGGATTTATC TGCTCTTCGC GTTGAAGAAG TACAAA.ATGT CATTAATGCT ATGCAGAAAA TCTTAGAGTG TCCCATCTGT CTGGAGTTGA TCMAGGAACC TGTCTCCACA AAGTGTGACC 31 j:j WO 96/3327 1 PCTIUS,96OS621

LI

II

/1 /1

I;

ACATATTTTG

GTCCTTTATG

AACTTGTTGA

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AATTGGGATC

ATCAAGAATT

CAAAAAAGGC

CCAGTAATAA

ATCAGGGTAG

GCTCATTACA

AGGCTGAATT

GGGCTGGAAG

ATCTGAATGC

CAGAGAATCC

35 AAGTTAATGA

GGGAGTCTGA

AATATTCTGG

TATGTAAAAG

TTGGGAAAAC

TAATTATAGG

AATTAAAGCG

CAGATTTGGC

AGAATGGTCA

CTATTCAGAA

AAACGAAAGC

ACAATTCAAA

ATGCGCTTGA

TTGATAGTTG

CAAATTTTGC

TAAGAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAGGACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGAA

TGATTTGAAC

TTCTGTTTCA

GCATGAGAAC

CTGTAATAAA

TAAGGAAACA

TGATCCCCTG

TAGAGATACT

GTGGTTTTCC

ATCAAATGCC

TTCTTCAGAG

TGAAAGAGTT

CTATCGGAAG

AGCATTTGTT

TAAAAGGAGA

AGTTCAAAAG

AGTGATGAAT

TGAGAAAAAT

TGAACCTATA

AG-CACCTAAA

ACTAGTAGTC

TTCTAGCAGT

ATGCTGAAAC

ATAACCAAAA

AA7LATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

AAGCAGCGGA

GAAGATACCG

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

TGTAATGATA

TGTGAGAGAA

GAAGATGTTC

AGAAGTGATG

AAAGTAGCTG

AAAATAGACT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCAG

ACTCCTGAAA

ATTACTAATA

CCTAACCCAA

AGCAGCAGTA

AAGAATAGGC

AGTAGAAATC

GAAGAGATAA

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAACCGTGC

GTCTCAGTGT

TACAACCTCA

TTAATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAGC

GGCGGACTCC

AAGAATGGAA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

TACTGGCCAG

CAGTAGAGAG

TCCCCAACTT

AGATAATACA

GCCTTCATCC

TGATAAATCA

GTGGTCATGA

TAGAATCACT

TAAGCAATAT

TGAGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGA-2CAT

CAAAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAAAT

AGACAGAATG

AAGGAGCCAA

CAGCACAGAA

TAAGCAGAAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAAAT

TGATCCTCAT

TAATATTGAA

AAGCCATGTA

AGAGCGTCCC

TGAGGATTTT

GGGAACTAAC

GAATAAAACA

CGAAAAAGAA

GGAACTCGAA

GTCTTCTACC

TAATTGTACT

GTACAACCAA

CCTTCACAGT

AGATTTAGTC

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

AACCTTGGAA

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

AAAAAGGTAG

CTGCCATGCT

AGCATTCAGA

TCACATGATG

GAGGTAGATG

GAGGCTTTAA

GACAAAATAT

ACTGAAAATC

CTCACA1&ATA

ATCAAGAAAG

CAAACGGAGC

AAAGGTGATT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GAATTGCAAA

ATGCCAGTCA

300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 GGCACAGCAG AALACCTACAA CTCATGGAAG GTAAAGAACC TGCAACTGGA GCCAAGAAGA 32 WO 96133271 WO 9/337 1PCT/US96/05(t21

GTAACAAGCC

AGTTAACAAA

TTGTCAATCC

CTAATAATGC

AAAGATCTGT

AGTATCTCGT

GTGAGTCAGT

AATAGAAATG

GA;LACAAGCA

AAGGTTTCAA

TGTGCAACAT

GAATGTGAAC

ACAGTTAATA

GCCAAATGTA

GAAACTGGAC

CCACTTTTTC

AACTTTGAGG

ACAGTGAGCA

AGCAATATTA

GGTTCCAGTG

GCTATGCTTA

AATTGTAAGC

ACAGATTTCT

GCATCTCAGG

GATACTAGTT

CAGAAAGGAG

TACCGAAGAG

GAGCTTCCCT

ACTAGGCATA

TCATTGAAGA

GAACATCACC

GAATTGGAAG

AAATGAACAG

TGCACCTGGT

TAGCCTTCCA

TGAAGACCCC

AGAGTAGCAG

TACTGGAAGT

GTGCAGCATT

ACACAGAAGG

TAGAAATGGA

AGCGCCAGTC

TCTCTGCCCA

AAAAGGAAGA

TCACTGCAGG

GTATCAAAGG

TCATTACTCC

CCATCAAGTC

AACATTCAAT

CAATTAGCCG

ATGAAGTAGG

ATGAAAACAT

GATTAGGGGT

ATCCTGAAAT

CTCCATATCT

TTTGTTCTGA

TTGCTGAAAA

AGCTTAGCAG

GGGCCAAGAA

GCTTCCAACA

GCACCGTTGC

ATAGCTTAAA

TTAGTGAGGA

ACTTGACTGC

ACAAGTAAAA

TCTTTTACTA

AGAGAAGAAA

AAAGATCTCA

TATTTCATTG

TAGCACTCTA

TGAAAACCCC

CTTTAAGTAT

AGAAAGTGAA

ATTTGCTCCG

CTCTGGGTCC

AAATCAAGGA

CTTTCCTGTG

AGGCTCTAGG

AAATAAACAT

ATTTGTTAAA

GTCACCTGAA

TAATAACATT

TTCCAGTACT

TCAAGCAGAA

TTTGCAACCT

AAAAAAGCAA

GATTTCAGAT

GACACCTGAT

TGACATTAAG

GAGTCCTAGC

ATTAGAGTCC

CTTGTTATTT

TACCGAGTGT

TGACTGCAGT

AACAAAATGT

AAATACAAAC

GACATGACAG

AGTGTTCAAA

AAGAAGAGAA

TGTTAAGTGG

GTACCTGGTA

GGGAAGGCAA

AAGGGACTAA

CCATTGGGAC

CTTGATGCTC

TTTTCAAATC

TTAAAGAAAC

AACk~ATGAGT

GTTGGTCAGA

TTTTGTCTAT

GGACTTTTAC

ACTAAATGTA

AGAGAAATGG

AGAGAAAATG

AATGAAGTGG

CTAGGTAGAA

GAGGTCTATA

GAATATGAAG

AACTTAGAAC

GACCTGTTAG

GAAAGTTCTG

CCTTTCACCC

TCAGAAGAGA

GGTAAAGTAA

CTGTCTAAGA

AACCAGGTAA

TCTGCTAGCT

ACCCAGGATC

CGATACTTTC

TACCAGTGAA

ACTAGAAAiCA

AGAAAGGGTT

CTGATTATGG

AAACAGAACC

TTCATGGTTG

ATGAAGTTAA

AGTATTTGCA

CAGGAAATGC

AAAGTCCAAA

CTAATATCAA

AAGATAAGCC

CATCTCAGTT

AAA.ACCCATA

AGAAAAATCT

GAAATGAGAA

TTTTTAAAGA

GCTCCAGTAT

ACAGAGGGCC

AACAA.AGTCT

AAGTAGTTCA

AGCCTATGGG

ATGATGGTGA

CTGTTTTTAG

ATACACATTT

ACTTATCTAG

ACAATATACC

ACACAGAGGA

TATTGGCAAA

TGTTTTCTTC

CTTTCTTGAT

CCAGAGCTGA

-fTAAAGAAT

GTTAAAGTGT

TTGCAAACTG

CACTCAGGAA

AAATAAATGT

TTCCAAAGAT

CCACAGTCGG

GAATACATTC

AGAAGAGGAA

AGTCACTTTT

GCCTC-TACAG

AGTTGATAAT

CAGAGGCAAC

TCGTATACCA

GCTAGAGGAA

CATTCCAAGT

AGCCAGCTCA

TAATGAAATA

AAAATTGAAT

TCCTGGAAGT

GACTGTTAAT

AAGTAGTCAT

AATAAAGGAA

CAAAAGCGTC

GGCTCAGGGT

TGAGGATGAA

TTCTCAGTCT

GAATTTATTA

GGCATCTCAG

ACAGTGCAGT

TGGTTCTTCC

2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 U IN U I WO 96/33271 PC'rUS96/05621

AAACAAATGA

TCAGATGATG

GATTCAAACT

TGCTCAGGGC

CATAACCTGA

GGGAGCCAGC

CTGCGAAATC

GAATACCCTA

GATAGTTCTA

CCATCATTAG

TACCCATCTC

TCTGGGCCAC

CCTTACCTGG

GACAGAGCCC

GTTCCCCAAT

ACTGCTGGGT

TCAACAGAAA

TTTATGCTCG

U GAAGAGACTA

AAATATTTTC

TCTATTAAAG

AATGGAAGAA

AGGGGGCTAG

U TGGATGGTAC

ACAGGTGTCC

CATGCAATTG

GTAGCACTCT

CACTACTGA

GGCATCAGTC

AAGAAAGAGG

TAGGTGAAGC

TATCCTCTCA

TAAAGCTCC1,

CTTCTAACAG

CAGAACAA.AG

TAAGCCAGAA

CCAGTAAAAA

ATGATAGGTG

AAGAGGAGCT

ACGATTTGAC

AATCTGGAAT

CAGAGTCAGC

TGAAAGTTGC

ATAATGCAAT

GGGTCAACAA

TGTACAAGTT

CTCATGTTGT

TAGGAATTGC

AAAGAAAAAT

ACCACCAAGG

AAATCTGTTG

AGCTGTGTGG

ACCCAATTGT

GGCAGATGTG

ACCAGTGCCA

TGAAAGCCAG

AACGGGCTTG

AGCATCTGGG

GAGTGACATT

GCAGGAAATG

CTACCCTTCC

CACATCAGAA

TCCAGAAGGC

TAAAGAACCA

GTACATGCAC

CATTAAGGTT

GGAAACATCT

CAGCCTCTTC

TCGTGTTGGC

AGAATCTGCC

GGAAGAAAGT

AAGAATGTCC

TGCCAGAAAA

TATGAAAACA

GGGAGGAAAA

GCTGAATGAG

TCCAAAGCGA

CTATGGGCCC

TGCTTCTGTG

GGTTGTGCAG

TGAGGCACCT

GGAGCTGGAC

GGAGTTGGTC

GAAGAAAATA

TGTGAGAGTG

TTAACCACTC

GCTGAACTAG

ATCATAAGTG

AAAGCAGTAT

CTTTCTGCTG

GGAGTGGAAA

AGTTGCTCTG

GTTGATGTGG

TACTTGCCAA

TCTGATGACC

AACATACCAT

CAGAGTCCAG

GTGAGCAGGG

ATGGTGGTGT

CACCACATCA

GATGCTGAGT

TGGGTAGTTA

CATGATTTTG

GCAAGAGAAT

TTCACCAACA

GTGAAGGAGC

CCAGATGCCT

GTGGTGACCC

ACCTACCTGA

TGAGTGACAA

ATCAAGAAGA

AAACAAGCGT

AGCAGAGGGA

AAGCTGTGTT

ACTCTTCTGC

TAACTTCACA

ACAAGTTTGA

GGTCATCCCC

GGAGTCTTCA

AGGAGCAACA

GGCAAGATCT

CTGAATCTGA

CTTCAACCTC

CTGCTGCTCA

AGAAGCCAGA

CTGGCCTGAC

CTTTAACTAA

TTGTGTGTGA

GCTATTTCTG

AAGTCAGAGG

CCCAGGACAG

TGCCCACAGA

TTTCATCATT

GGACAGAGGA

GAGAGTGGGT

TACCCCAGAT

GGAATTGGTT

GCAZ.AGCATG

CTCTGAAGAC

TACCATGCAA

AGAACAGCAT

CCTTGAGGAC

GAAAAGTAGT

GGTGTCTGCA.

TTCTAAATGC

GAATAGAAAC

GCTGGAAGAG

AGAGGGAACC

TCCTTCTGAA

TGCATTGAAA

TACTACTGAT

ATTGACAGCT

CCCAGAAGAA

TCTAATTACT

ACGGACACTG

GGTGACCCI.G

AGATGTGGTC

AAAGATCTTC

TCAACTGGAA

CACCCTTGGC

CAATGGCTTC

GTTGGACAGT

CCCCCACAGC

4140 4200 4260 4320 4380 4440 -4500 4560 4620 A.68 0 4740 4800 4860 4920 4900 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5709 INFORMATION FOR SEQ ID NO:8: SEQUENCE CHARACTERISTICS: LENGTH: 5709 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear

IUI~~

WO 96/33271 PCT[US96/05621 (ii) MOLECULE TYPE: CDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTIGTGGG

CCGGCC

TGGATTTATC

TCTTAGAGTG

ACATATTTTG

GTCCTTTATG

AACTTGTTGA

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AATTGGGATC

ATCAAGAATT

CAAAAAAGGC

CCAGTA.ATAA

ATCAGGGTAG

GCTCATTACA

AGGCTGAATT

GGGCTGGAAG

ATCTGAATGC

CAGAGAATCC

AAGTTAATGA

so GGGAGTCTGA

AATATTCTGG

TATGTAAAAG

TTGGGAAAAC

TAATTATAGG

AATTAAAGCG

CAGATTTGGC

AGAATGGTCA

GGAGGCCTTC

TGCTCTTCGC

TCCCATCTGT

CAAATTTTGC

TAAGAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAGGACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGAA

TGATTTGAAC

TTCTGTTTCA

GCATGAGALAC

CTGTAATAAA

TAAGGAAACA

TGATCCCCTG

TAGAGATACT

GTGGTTTTCC

ATCAAATGCC

TTCTTCAGAG

TGAAAGAGTT

CTATCGGAAG

AGCATTTGTT

TAAAAGGAGA

AGTTCAkP.AG

AGTGATGAAT

ACCCTCTGCT

GTTGAAGAAG

CTGGAGTTGA

ATGCTGAAAC

ATAACCAAAA

AAAATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

A.AGCAGCGGA

GAAGATACCG

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

TGTAATGATA

TGTGAGAGAA

GAAGATGTTC

AGAAGTGATG

AAAGTAGCTG

AAAATAGACT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCAG

ACTCCTGAA

ATTACTAATA

CTGGGTAAAG

TACAAAATGT

TCAAGGAACC

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAACCGTGC

GTCTCAGTGT

TACAACCTCA

TTAATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAGC

GGCGGACTCC

AAGAATGGA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

TACTGGCCAG

CAGTAGAGAG

TCCCCAACTT

AGATAATACA

GCCTTCATCC

TGATAAATCA

GTGGTCATGA

GTTTCTCAGA

TTCATTGGAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGAACAT

CAAAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAAAT

AGACAGAATG

AAGGAGCCAA

CAGCACAGAA

TAAGCAGAAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAAAT

TGATCCTCAT

TAATATTGAA

AAGCCATGTA

AGAGCGTCCC

TGAGGATTTT

GGGAACTXAC

GAATAAAACA

TAACTGGGCC

CAGAAAGAAA

ATGCAGAAAA

AAGTGTGACC

CCTTCACAGT

AGATTTAGTC

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

AACCTTGGAA

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

AAAAAGGTAG

CTGCCATGCT

AGCATTCAGA

TCACATGATG

GAGGTAGATG

GAGGCTTTAA

GACAAAATAT

ACTGAAAA'rC

CTCACAAATA

ATCAAGAAAG

CAAACGGAGC

AAAGGTGATT

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 *1 22 WO 96/33271 PCTIUS96/05621

CTATTCAGAA

AAACGAAAGC

ACAATTCAAA

ATGCGCTTGA

TTGATAGTTG

GGCACAGCAG

GTAACAAGCC

AGTTAACAAA

TTGTCAATCC

CTAATAATGC

AAAGATCTGT

AA.AGTATCTC

GTGTGAGTC]

ATAATAGAAA

GGGAAACAAG

TCAAGGTTTC

AATGTGCAAC

GAATGTGAAC

ACAGTTAATA

GCCAAATGTA

GAAACTGGAC

CCACTTTTTC

AACTTTGAGG

ACAGTGAGCA

AGCAATATTA

GGTTCCAGTG

GCTATGCTTA

AATTGTAAGC

ACAGATTTCT

GCATCTCAGG

GATACTAGTT

CAGAAAGGAG

TGAGAAAAAT

TGAACCTATA

AGCACCTAAA

ACTAGTAGTC

TTCTAGCAGT

AAACCTACAA

AAATGAACAG

TGCACCTGGT

TAGCCTTCCA

TGAAGACCCC

AGAGAGTAGC

GTTACT(.. "AA

GTGTGCAGCA

TGACACAGAA

CATAGAAATG

AAAGCGCCAG

ATTCTCTGCC

AAAAGGAAGA

TCACTGCAGG

GTATCAAAGG

TCATTACTCC

CCATCAAGTC

AACATTCAAT

CAATTAGCCG

ATGAAGTAGG

ATGAAAACAT

GATTAGGGGT

ATCCTGAAAT

CTCCATATCT

TTTGTTCTGA

TTGCTGAAAA

AGCTTAGCAG

CCTAACCCAA

AGCAGCAGTA

AAGAATAGGC

AGTAGAkATC

GAAGAGATAA

CTCATGGAAG

ACAAGTAAAA

TCTTTTACTA

AGAGAAGAAA

AAAGATCTCA

AGTATTTCAT

GTTAr-_ -C

TTTGAAAACC

GGCTTTAAGT

GAAGAAAGTG

TCATTTGCTC

CACTCTGGGT

AAATCAAGGA

CTTTCCTGTG

AGGCTCTAGG

AA.ATAAACAT

ATTTGTTAAA

GTCACCTGAA

TAATAACATT

TTCCAGTACT

TCAAGCAGAA

TTTGCAACCT

AAAAAAGCAA

GATTTCAGAT

GACACCTGAT

TGACATTAAG

GAGTCCTAGC

TAGAATCACT

TAAGCAATAT

TGAGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GTAAAGAACC

GACATGACAG

AGTGTTCAAA

AAGAAGAGAA

TGTTAAGTGG

TGGTACCTGG

TAGGGAAGGC

CCAAGGGACT

ATCCATTGGG

AACTTGATGC

CGTTTTCAAA

CCTTAAAGAC

AAGAATGAGT

GTTGGTCAGA

TTTTGTCTAT

GGACTTTTAC

ACTAAATGTA

AGAGAAATGG

AGAGAAAATG

AATGAAGTGG

CTAGGTAGAA

GAGGTCTATA

GAATATGAAG

AACTTAGAAC

GACCTGTTAG

GAAAGTTCTG

CCTTTCACCC

CGAAAAAGAA

GGAACTCGAA

GTCTTCTACC

TAATTGTACT

GTACAACCAA

TGCAACTGGA

CGATACTTTC

TACCAG"'GAA

ACTAGAAACA

AGAAAGGGTT

TACTGATTAT

AAAAACAGAA

AATTCATGGT

ACATGAAGTT

TCAGTATTTG

TCCAGGAAAT

AAAGTCCAAA

CTAATATCAA

AAGATAAGCC

CATCTCAGTT

AAAACCCATA

AGAAAAATCT

GAAATGAGAA

TTTTTAAAGA

GCTCCAGTAT

ACAGAGGGCC

AACAAAGTCT

AAGTAGTTCA

AGCCTATGGG

ATGATGGTGA

CTGTTTTTAG

ATACACATTT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GAATTGCAA

ATGCCAGTCA

GCCAAGAAGA

CCAGAGCTGA

CTTAAAGAAT

GTTAAAGTGT

TTGCAAACTG

GGCACTCAGG

CCAAATAAAT

TGTTCCAAAG

AACCACAGTC

CAGAATACAT

GCAGAAGAGG

AGTCACTTTT

GCCTGTACAG

AGTTGATAAT

CAGAGGCAAC

TCGTATACCA

GCTAGAGGAA

CATTCCAAGT

AGCCAGCTCA

TAATGAAATA

AAAATTGAAT

TCCTGGAAGT

GACTGTTAP.,T

AAGTAGTCAT

AATAAAGGAA

CAAAAGCGTC

GGCTCAGGGT

1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3400 3540 3600 3660 3720 tvl-w UUUAAATAGGAA GATACTAGTT TTGCTGAAA 3600 I I I

I,

WO 96/33271 PCTTJS96/05621

TACCGAAGAG

GAGCTTCCCT

ACTAGGCA'

TCATTGAAGA

GAACATCACC

GAATTGGAAG

AAACAAATGA

TCAGATGATG

GATTCAAACT

TGCTCAGGGC

CATAACCTGA

GGGAGCCAGC

CTGCGAAATC

GAATACCCTA

GATAGTTCTA

CCATCATTAG

TACCCATCTC

TCTGGGCCAC

CCTTACCTGG

GACAGAGCCC

GTTCCCCAAT

ACTGCTGGGT

TCAACAGAAA

TTTATGCTCG

GAAGAGACTA

AAATATTTTC

TCTATTAAAG

AATGGAAGAA

AGGGGGCTAG

TGGATGGTAC

ACAGGTGTCC

CATGCAATTG

GGGCCAAGAA

GCTTCCAACA

CGTTGC

ATAGCTTAAA

TTAGTGAGGA

ACTTGACTGC

GGCATCAGTC

AAGAAAGAGG

TAGGTGAAGC

TATCCTCTCA

TAAAGCTCCA

CTTCTAACAG

CAGAACAAAG

TAAGCCAGAA

CCAGTAAAAA

ATGATAGGTG

AAGAGGAGCT

ACGATTTGAC

AATCTGGAAT

CAGAGTCAGC

TGAAAGTTGC

ATAATGCAAT

GGGTCAACAA

TGTACAAGTT

CTCATGTTGT

TAGGAATTGC

AALAGAAAAAT

ACCACCAAGG

AAATCTGTTG

AGCTGTGTGG

ACCCAATTGT

GGCAGATGTG

ATTAGAGT.CC

CTTGTTATTT

TACCGAGTGT

TGACTGCAGT

AACAAAATGT

AAATACAAAC

TGAAAGCCAG

AACGGGCTTG

AGCATCTGGG

GAGTGACATT

GCAGGA.AATG

CTACCCTTCC

CACATCAGAA

TCCAGAAGGC

TAAAGAACCA

GTACATGCAC

CATTAAGGTT

GGAAACATCT

CAGCCTCTTC

TCGTGTTGGC

AGAATCTGCC

GGAAGAAAGT

AAGAATGTCC

TGCCAGAAAA

TATGAAAACA

GGGAGGAAAA

GCTGAATGAG

TCCAAAGCGA

CTATGGGCCC

TGCTTCTGTG

GGTTGTGCAG

TGAGGCACCT

TCAGAAGAGA

GGTAAAGTAA

CTGTCTAAGA

AACCAGGTAA

TCTGCTAGCT

ACCCAGGATC

GGAGTTGGTC

GAAGAAAATA

TGTGAGAGTG

TTAACCACTC

GCTGAACTAG

ATCATAAGTG

AAAGCAGTAT

CTTTCTGCTG

GGAGTGGAAA

AGTTGCTCTG

GTTGATGTGG

TACTTGCCAA

TCTGATGACC

AACATACCAT

CAGAGTCCAG

GTGAGCAGGG

ATGGTGGTGT

CACCACATCA

GATGCTGAGT

TGGGT.AGTTA

CATGATTTTG

GCAAGAGAAT

TTCACCAACA

GTGAAGGAGC

CCAGATGCCT

GTGGTGACCC

ACTTATCTAG

ACAATATACC

ACACAGAGGA

TATTGGCAAA

TGTTTTCTTC

CTTTCTTGAT

TGAGTGACAA

ATCAAGAAGA

AAACAAGCGT

AGCAGAGGGA

AAGCTGTGTT

ACTCTTCTGC

TAACTTCACA

ACAAGTTTGA

GGTCATCCCC

GGAGTCTTCA

AGGAGCAACA

GGCAAGATCT

CTGAATCTGA

CTTCAACCTC

CTGCTGCTCA

AGAAGCCAGA

CTGGCCTGAC

CTTTAACTAA

TTGTGTGTGA

GCTATTTCTG

AAGTCAGAGG

CCCAGGACAG

TGCCCACAGA

TTTCATCATT

GGACAGAGGA

GAGAGTGGGT

TGAGGATGAA

TTCTCAGTCT

GAATTTATTA

GGCA'2'CTCAG

ACAGTGCAGT

TGGTTCTTCC

GGAATTGGTT

GCAAAGCATG

CTCTGAAGAC

TACCATGCX-A

AGAACAGCAT

CCTTGAGGAC

GAAAAGTAGT

GGTGTCTGCA

TTCTAAATGC

GAATAGAAAC

GCTGGAAGAG

AGAGGGAACC

TCCTTCTGAA

TGCATTGAAA

TACTACTGAT

ATTGACAGCT

CCCAGAAGAA

TCTAATTACT

ACGGACACTG

GGTGACCCAG

AGATGTGGTC

AAAGATCTTC

TCA .CTGGAA

CACCCTTGGC

CAATGGCTTC

GTTGGACAGT

3780 3840 3900 3960 4020 408) 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 WO 96/33271 WO 9633271PCT[US96/05621 GTAGCACTCT ACCAGTGCCA GGAGCTGGAC P ;I.,ACCTGA TACCCCAGAT CCCCCACAGC

CACTACTGA

INFORM.ATION FOR SEQ ID NO:9: SEQUENCE CHARACTZRISTICS: LENGTH: 5709 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: 5700 5709 AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG

CCTGCGCTCA

TGGA'1-I ATC

TCTTAGAGTG

ACATATTTTG

GTCCTTTATG

AACTTGTTGA

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AATTGGGATC

ATCAAGAATT

CAAAAAAGGC

CCAGTAATAA

ATCAGGGTAG

GCTCATTACA

AGGCTGAATT

GGGCTGGAAG

ATCTGAATGC

CAGAGAATCC

AAGTTAATGA

GGGAGTCTGA

AATATTCTGG

GGAGGCCTTC

TGCTCTTCGC

TCCCATCTGT

CAAATTTTGC

T %GAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAC-GACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGAA

TGATTTGAAC

TTCTGTTTCA

GCATGAGAAC

CTGTAATAAA

TAAGGAAACA

TGATCCCCTG

TAGAGATACT

GTGGTTTTCC

ATCAAATGCC

TTCTTCAGAG

ACCCTCTGCT

GTTGAAGAAG

CTGGAGTTGA

ATGCTGAAAC

ATAACCAAAA

AAAATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

AAGCAGCGGA

GAAGATACCG

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

TGTAATGATA

TGTGAGAGAA

GAAGATGTTC

AGAAGTGATG

AAAGTAGCTG

AAAATAGACT

CTGGGTAAAG

TACAAAATGT

TCAAGGAACC

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAACCGTGC

GTCTCAGTGT

TACAACCTCA

TTAATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAUC

GGCGGACTCC

AAGAATGGAA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

TACTGGCCAG

GTTTCTCAGA

TTCATTGGAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGAACAT

CAAAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAAAT

AGACAGAATG

AAGGAGCCAA

CAGCACAGAA

TAA( i J.-AAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAAAT

TGATCCTCAT

TAACTGGGCC

CAGAAAGAAA

ATGCAGAAAA

AAGTGTGACC

CCTTCACAGT

AGATTTAGTC

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

AACCTTGGAA

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

AAAAAGGTAG

CTGCCATGCT

AG3CATTCAGA

TCACATGATG

GAGGTAGATG

GAGGCTTTAA

6u 120 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 PCT[US96/05621 'WO 96/3327 1

TATGTAAAAG

TTGGGAAAAC

T-iATTATAGG

AATTAAAGCG

CAGATTTGGC

AGAATGGTCA

CTATTCAGAA

AAACGAAAGC

ACAATTCAAA

ATGCGCTTGA

TTGATAGTTG

GGCZACAGCAG

GTAACAAGCC

AGTTAAC UAA

TTGTCAAICC

CTAATAATCJ'C

AAAGATCTGT

AAAGTATCTC

GTdTGAGTCA

ATAATAGAAA

GGGAAAWAAG

TCAAGGTTTC

AATCTGCAAC

TTGAATGTGA

ACAGTTAATA

GCCAAATGTA

GAAACTGGAC-

CCACTTTTTC

AACTTTGAGG

ACAGTGAGCA

AGCAATATTA

GGTTCCAGTG

TGAAAGAGTT

CTATCGGAAG

AGCATTTGTT

TAAAAGGiGA

AGTTCAAAAG

AGTGA'rGAAT

TGAGAAAAAT

'rGAACCTATA

AGCACCTAAA

ACTAGTAGTC

TTCTAGCAGT

AAACCTACAA

A,;ATGAACAG

TGCACCTGGT

TAGCCTTCCA

TGAAGACCCC

P -AG~ C

GTTACTGGAA

GTGTGCAGCA

TGACACAGAA

CATAGAAATG

AAAGCGCCAG

ATTCTCTGCC

ACAAAAG6,AA

TCACTGCAGG

GTATCAAAGG

TCATTACTCC

CCATCAAGTC

AACATTCAAT

Ck..TTAGCCG

ATGAAGTAGG

ATGAAAACAT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCAG

ACTCCTGAAA

ATTACTAATA

CCTAACCCAA

AGCAGCAGTA

AAGAATAGGC

AGTAGAAATC

GAAGAGATAA

CTCATGGAAG

ACAAGT .'Ak TCTTTTACTh

AGAGAAGAA.A

AA.AGATCTCA

AGTATTTCAT

GTTAGCACTC

TTTGAAAACC

GGCTTTAAGT

GAAGAPAGTG

TCATTTGCTC

CACTCTGGGT

GAAAATCAAG

CTTTCCTGTG

AG(",CTCTAGG

AAATAAACAT

ATTTGTTAAA

GTCACCTGAA

TAAP.ATACATT

TTCCAGTACT

TCAAGCAGAA

CAGTAGAGAG

TCCCC AACTT

AGATAATACA

GCCTTCATCC

TGATAAATCA

GTGGTCATGA

TAGAATCACT

TAAGCAATAT

TG AGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GTAAAGAACC

GACATGACAG

AGTGTTCAAA

AAGAAGAGAA

TGTTPAAGTGG

TGGTACCTGG

TAGGGAAGGC

CCAAGGGACT

ATCCATTGGG

AACTTGATGC

CGTTTTCAAA

CCTTAUA.GAA

GAAAGAATGA

GTTGGTCAGA

TTTTGTCTAT

GGACTTTTAC

ACTAAATGTA

AGAGAAATG

AGAGAAAATG

AATGAAGTGG

CTAGGTAGAA

TAATATTGAA

AAGCCATGTA

AGAGCOTCCC

TGAGGATTIT

GGGAACTAAC

GAATAAAACA

CGAAAAAGAA

GGAACTCGAA

GTCTTCTACC

TA) .TGTACT

GTACAACCAA

TGCAACTGGA

CGATACTTTC

TACCAGTGAA

ACTAGAAACA

AGAAAGGGTT

TACTGNTTAT

AAAAACAGAA

\ATTCATGGT

AC ATCAAGTT TCA('. TATTTG TCCAGGAAATr

ACAAAGTCCA

GTAATATCAA

AAGATAAGCC

CATCTC'hGTT

AAAACCCATA

AGPAkAAATCT

GAAATGAGAA

TTTTTAAAGA

GCTCCAGTAT

ACAGAGGGCC

GACAAAATAT

ACTGAAAATC

CTCACAAATA

ATCAAGAAAG

CAAACGGAGC

A21LAGGTGATT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GA'TGCAAA

A±i ,AGTCA

GCCAAGAAGA

CCAGAGCTGA

CTTAAAGAAT

GTTAAAGTGT

TTGCAAACTG

GGCACTCAGG

CC-AAATAAAT

TGTTCCAAAG

AACCACAGTC

CAGAATACAT

GCAGAAGAGG

AAAGTCACTT

GC~mGTACAG

AGTTGATAAT

CAGAGGCAAC

TCGTATACCA

GCTAGAGGAA

CATTCCAAGT

AGCCAGCTCA

TAATGAAATA

AAAATTGAAT

1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2 700 2760 2820 2R80 2940 3000 3060 3120 318u 3240 3300 3360 11

L

WO 96/33271 FTU9152 FCT[US96/05621

GCTATGCTTA

AATTGTAAGC

ACAGATTTCT

GCATCTCAGG

GATACTAGTT

CAGAAAGGAG

TACCGAAGAG

GAGCTTCCCT

ACTAGGCATA

TCATTGAAGA

GAACATCACC

GAATTGGAAG

AAACAAATGA

TCAGATGATG

GATTCAAACT

TGCTCAGGGC

CATAACCTGA

GGGAGCCAGC

CTGCGAAATC

GAATACCCTA

GATAGTTCTA

CCATCATTAG

TACCCATCTC

TCTGGGCCAC

CCTTACCTGG

GACAGAGCCC

GTTCCCCAAT

ACTGCTGGGT

TCAACAGAAA

TTTATGCTCG

GAAGAGACTA

AAATATTTTC

GATTAGGGGT

ATCCTGAAAT

CTCCATATCT

TT TTCTGA

TT'GCTGAAAA

A:7CTTAGCAG

GGGCCAAGAA

GCTTCCAACA

GCACCGTTGC

ATAGCTTAAA

TTAGTGAGGA

ACTTGACTGC

GGCATCAGTC

AAGAAAGAGG

TAGGTGAAGC

TATCCTCTCA

TAAAGCTCCA

CTTCTAACAG

CAGAACAAAG

TAAGCCAGAA

CCAGTAAAAA

ATGATAGGTG

AAGAGGAGCT

ACGATTTGAC

AATCTGGAAT

CAGAGTCAGC

TGAAAGTTGC

ATAATGCAAT

GGGTCAACAA

TGTACAAGTT

CTCATGTTGT

TAGGAATTGC

TTTGCAACCT

AAAAAAGCAA

GATTTCAGAT

GACACCTGAT

TGACATTAAG

GAGTCCTAGC

ATTAGAGTCC

CTTGTTATTT

TACCGAGTGT

TGACTGCAGT

AACAAAATGT

AAATACAAAC

TGAAAGCCAG

AACGGGCTTG

AGCATCTGGG

GAGTGACATT

GCAGGAAATG

CTACCCTTCC

CACATCAGAA

TCCAGAAGGC

TAAAGAACCA

GTACATGCAC

CATTYAAGGTT

GGAAACATCT

CAGCCTCTTC

TCGTGTTGGC

AGAATCTGCC

GGAAGAAAGT

AAGAATGTCC

TGCCAGAAAA

TATGAAAACA

GGGAGGAAAA

GAGGTCTATA

GAATATGAAG

AACTTAGAAC

GACCTGTTAG

GAAAGTTCTG

CCTTTCACCC

TCAGAAGAGA

GGTAAAGTAA

CTGTCTAAGA

AACCAGGTAA

TCTGCTAGCT

ACCCAGGATC

GGAGTTGGTC

GAAGAAAATA

TGTGAGAGTG

TTAACCACTC

GCTGAACTAG

ATCATAAGTG

AAAGCAGTAT

CTTTCTGCTG

GGAGTGGAAA

AGTTGCTCTG

GTTGATGTGG

TACTTGCCAA

TCTGATGACC

AACATACCAT

CACAGTCCAG

GTGAGCAGGG

ATGGTGGTGT

CACCACATCA

GATGCTGAGT

TGGGTAGTTA

AACAAAGTCT

AA.GTAGTTCA

AGCCTATGGG

ATGATGGTGA

CTGTTTTXTAG

ATACACATTT

ACTTATC'TAG

ACAATATACC

ACACAGAGGA

TATTGGCAAA

TGTTTTCTTC

CTTTCTTGAT

TGAGTGACAA

ATCAAGAAGA

AAACAAGCGT

AGCAGAGGGA

AAGCTGTGTT

ACTCTTCTGC

TAiACTTCACA

ACAAGTTTGA

GGTCATCCCC

GGAGTCTTCA

AGGAGCAACA

GGCAAGATCT

CTGAATCTGA

C!TTCAACCTC

CTGCTGCTCA

AGAAGCCAGA

CTGGCCTGAC

CTTTAACTA.

TTGTGTGTGA

GCTATTTCTG

TCCTGGAAGT

GACTGTTAAT

AAGTAGTCAT

AATAAAGGAA

CAAAAGCGTC

GGCTCAGGGT

TGAGGATGAA

TTCTCAGTCT

GAATTTATTA

GGCATCTCAG

ACAGTGCAGT

TGGTTCTTCC

GGAATTGGTT

GCAAAGCATG

CTCTGAAGAC

TACCATGCAA

AGAACAGCAT

CCTTGAGGAC

GAAAAGTAGT

GGTGTCTGCA

TTCTAAATGC

GAATAGAAAC

GCTGGAAGAG

AGAGGGAACC

TCCTTCTGAA

TGCATTGAAA

TACTACTGAT

ATTGACAGCT

CCCAGAAGAA

TCTAATTACT

ACGGACACTG

GGTGACCCAG

3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 27 Ii WO 96133271 TCTATTAAAG AAAGAA.A LI AATGGAAGAA ACCACCAA AGGGGGCTAG AAATCTGI TGGATGGTAC AGCTGTGT CATGCAATTG GGCAGATG GACATTC ACCATI

EG

TYPE:

STPAW

TOPOL

(ii) MOLECULE TI (xi) SEQUENCE D

I

PCTUS96105621 AT GCTGAATGAG CATGATYT'VU GG TCCAAAGCGA GCAAGAGAAT 'TG CTATGGGCCC TTCACCAACA 'GG TGCTTCTGTG GTGAAGGAGC 'GT GGTTGTGCAG CCAGATGCCT TG TGAGGCACCT GTGGTGACCC CA GGAGCTGGAC ACCTACCTGA SEQ ID NO:lO:

HARACTERISTICS:

5711 base pairs nucleic acid (DEDNESS: double ,OGY: linear 'YPE: cDNA )ESCRIPTION: SEQ ID NO:1O:

AAGTCAGAGG

CCCAGGACAG

I'GCCCACAGA

TTTCATCATT

GGACAGAGGA

GAGAGTGGGT

TACCCCAGAT

AGATGTGGTC

AAAGATCTTC

TCAACTGGAA

CACCCTTGGC

CAATGGCTTC

GTTGGACAGT

CCCCCACAGC

5340 5400 5460 5520 5580 5640 5700 5709 AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG GTTTCTCAGA TAACTGGGCC Af

CCTGCGCTCA

TGGATTTATC

TCTTAGAGTG

ACATATTTTG

GTCCTTTATG

AACTTGTTGA

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AATTGGGATC

ATCAAGAATT

CAAAAAAGGC

CCAGTAATAA

ATCAGGGTAG

GCTCATTACA

AGGCTGAATT

GGAGGCCTTC

TGCTCTTCGC

TCCCATCTGT

CAAATTTTGC

TAAGAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAGGACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGAA.

TGATTTGAAC

TTCTGTTTCA

GCATGAGAAC

CTGTAATAAA

ACCCTCTGCT

GTTGAAGAAG

CTGGAGTTGA

ATGCTGAAAC

ATAACCAAAA

AAAATCATTT

GCAAAAAkAGG

ATGGGCTACA

CAGGAAACCA

AAGCAGCGGA

GAAGATAC.

ACCCCTCAAG

TTTTCTGAG.A

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

CTGGGTAAAG

TACAAAATGT

TCAAGGAACC

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAACCGTGC

GTCTCAGTGT

TACAACCTCA

TTA.ATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAGC

TTCATTGGAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGAACAT

CAANAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAAAT

AGACAGAATG

AAGGAGCCAA

CAGAAAGAAA

ATGCAGAAAA

AAGTGTGACC

CCTTCACAGT

AGATTTAGTC

GGTTTGGAGT

CTAA.AAGATG

CTACAGAGTG

AACCTTr-CAA

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 WO 96/33271 WO 9633271PCT[US96/05621

GGGCTGGAAG

ATCTGAATGC

CAGAGAATCC

AAGTTAATGA

GGGAG-TCTGA

AATATTCTGG

TATGTAAA.AG

TTGGGAAAAC

TAATTATAGG

AATTAAAGCG

CAGATTTGGC

AGAATGGTCA

CTATTCAGAA

A.ACGAAAGC

ACAATTCAAA

ATGCGCTTGA

TTGATAGTTG

GGCACAGCAG

GTAACAAGCC

AGTTAACAAA

TTGTCAATCC

CTAATAATGC

AAAGATCTGT

AAAGTATCTC

GTGTGAGTCA

ATAATAGAAA

GGGAAACAAG

TCAAGGTTTC

AATGTGCAAC

TTGAATGTGA

AGACAGTTAA

ATGCCAAATG

TAAGGAAACA TGTAATGATA GGCGGACTCC CAGCACAGAA AAAAAGGTAG TGATCCCCTG TGTGAGAGAA

TAGAGATACT

GTGGTTTTCC

ATCAAATGCC

TTCTTCAGAG

TGAAAGAGTT

CTATCGGAAG

AGCATTTGTT

TAAAAGGAGA

AGTTCAAAAG

AGTGATGAAT

TGAGAAAAAT

TGAACCTATA

AGCACCTAAA

ACTAGTAGTC

TTCTAGCAG-T

AAACCTACAA

AAATGAACAG

TGCACCTGGT

TAGCCTTCCA

TGAAGACCCC

AGAGAGTAGC

GTTACTGGAA

GTGTGCAGCA

TGACACAGAA

CATAGAAATG

AAAGCGCCAG

ATTCTCTGCC

ACAAAAGGAA

TATCACTGCA

ThGTATCAAPA

GAAGATGTTC

AGAAGTGATG

AAAGTAGCTG

AAAATAGACT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCIG

ACTCCTGAAA

ATTACTAATA

CCTAACCCAA

AGCAGCAGTA

AAGAATAGG,

AGTAGAAATC

GAAGAGATAA

CTCATGGAAG

ACAAGTAAAA

TCTTTTACTA

AGAGAAGAAA

AAAGATCTCA

AGTATTTCAT

GTTAGCACTC

TTTGAAAACC

GGCTTTAAGT

GAAGAAAGTG

TCATTTGCTC

CACTCTGGGT

GAAAATCAAG

GGCTTTCCTG

GGAGGCTCTA

AAGAATGGAA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

TACTGGCCAG

CAGTAGAGAG

TCCCCAACTT

AGATAATACA

GCCTTCATCC

TGATAA-ACA

GTGGTCATGA

TAGAATCACT

TAAGCAATAT

TGAGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GTAAAGAACC

GACATGACAG

AGTGTTCAAA

AAGAAGAGAA

TGTTAAGTGG

TGGTACCTGG

TAGGGAAGGC

CCAAGGGACT

ATCCATTGGG

AACTTGATGC

CGTTTTCAAA

CCTTAAAGAA

GAAAGAATGA

TGGTTGGTCA

GGTTTTGTCT

TAAGCAGAAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAAAT

TGATCCTCAT

TAATATTGAA

AAGCCATGTA

AGAGCGTCCC

TGAGGATTTT

GGGAACTAAC

GAATAAAACA

CGAAAAAGAA

GGAACTCGAA

GTCTTCTACC

TAATTGTACT

GTACAACCAA

TGCAACTGGA

CGATACTTTC

TACCAGTGAA

ACTAGAAACA

AGAAAGGGTT

TACTGATTAT

AAAAACAGAA

AATTCATGGT

ACATGAAGTT

TCAGTATTTG

TCCAGGAAAT

ACAAAGTCCA

GTCTAATATC

GAAAGAT

7

AAG

ATCATCTCAG

CTGCCAkTGCT

AGCATTCAGA

TCACATGATG

GAGGTAGATG

GAGGCTTTAA

GACAAAATAT

A.TGAAAATC

CTCACAAATA

ATCAAPGAAAG

CAAACGGAGC

AAAGGTGATT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GAATTGCAAA

ATGCCAGTCA

GCCAAGAAGA

CCAGAGCTGA

CTTA.AAGAAT

GTTAAAGTGT

TTGCAAACTG

GGCACTCAGG

CCAAATAAAT

TGTTCCAAAG

AACCACAGTC

CAGAATACAT

GCAGAAGAGG

AAAGTCACTT

AAGCCTGTAC

CCAGTTGATA

TTCAGAGGCA

1140 1200 1260 1320 -380 1440 -1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 WO 96/33271 WO 9633211PCTIIJS96/0S621

ACGAAACTGG

CACCACTTTT

AAAACTTTGA

GTACAGTGAG

CAAGCAATAT

TAGGTTCCAG

ATGCTATGCT

GTAATTGTAA

ATACAGATTT

ATGCATCTCA

AAGATACTAG

TCCAGAAAGG

GTTACTGAAG

AAGAGCTTCC

CTACTAGGCA

TATCATTGAA

AGGAACATCA

GTGAATTGGA

CCAAACAAAT

TTTCAGATGA

TGGATTCAAA

ACTGCTCAGG

AACA'?AACCT

ATGGGAGCCA

ACCTGCGAAA

GTGAATACCC

CAGATAGTTC

GCCCATCATT

ACTACCCATC

AGTCTGGGCC

CCCCTTACCT

ACTCATTACT

TCCCATCAAG

GGAACATTCA

CACAATTAGC

TAATGAAGTA

TGATGAAAAC

TAGATTAGGG

GCATCCTGAA

CTCTCCATAT

GGTTTGTTCT

TTTTGCTGAA

AGAGCTTPLGC

AGGGGCCAAG

CTGCTTCCAA

TAGCACCGTT

GAATAGCTTA

CCTTAGTGAG

AGACTTGACT

GAGGCATCAG

TGAAGAAAGA

CTTAGGTGAA

GCTATCCTCT

GATAAAGCTC

GCCTTCTAAC

TCCAGAACAA

TATAAGCCAG

TACCAGTAAA

AGATGATAGG

TCAAGAGGAG

ACACGATTTG

GGAATCTGGA

CCAAATAAAC

TCATTTGTTA

ATGTCACCTG

CGTAATAACA

GGTTCCAGTA

ATTCAAGCAG

GTTTTGCAAC

ATAAAAAAGC

CTGATTTCAG

GAGACACCTG

AATGACATTA

AGGAGTCCTA

AAATTAGAGT

CACTTGTTAT

GCTACCGAGT

AATGACTGCA

GAAACAAAAT

GCAAATACAA

TCTGAAAGCC

GGAACGGGCT

GCAGCATCTG

CAGAGTGACA

CAGCAGGAAA

AGCTACCCTT

AGCACATCAG

AATCCAGAAG

AATAAAGAAC

TGGTAkCATGC CTCAkTTAAGG

ACGGAAACAT

ATCAGCCTCT

ATGGACTTTT

AAACTAAATG

AAAGAGAAAT

TTAGAGAAAA

CTAATGAAGT

AACTAGGTAG

CTGAGGTCTA

AAGAATATGA

ATAACTTAGA

ATGACCTGTT

AGGAAAGTTC

GCCCTTTCAC

CCTCAGAAGA

TTGGTAAAGT

GTCTGTCTAA

GTAACCAGGT

GTTCTGCTAG

ACACCCAGGA

AGGGAGTTGG

TGGAAGAAA1A

GGTGTGAGAG

TTTTAACCAC

TGGCTGAACT

CCATCATAAG

AAAAAGCAGT

GCCTTTCTGC

CAGGAGTGGA

ACAGTTGCTC

TTG'TTGATGT

CTTACTTGCC

TCTCTGATGA

ACAAA.ACCCA

TAAGAAAAAT

GGGAAATGAG

TGTTTTTAAA

GGGCTCCAGT

AAACAGAGGG

TAAACAAAGT

AGAAGTAGTT

ACAGCCTATG

AGATGATGGT

TGCTGTTTTT

CCATACACAT

GAACTTATCT

AAACAATATA

GAACACAGAG

AATATTGGCA

CTTGTTTTCT

TCCTTTCTTG

TCTGAGTGAC

TAATCAAGAA

TGAAACAAGC

TCAGCAGAGG

AGAAGCTGTG

TGACTCTTCT

ATTAACTTCA

TGACAkAGTTT

AAGGTCATCC

TGGGAGTCTT

GGAGGAGCAA

AAGGCAAGAT

CCCTGAATCT

TATCGTATAC

CTGCTAGAGG

AACATTCCAA

GAAGCCAGCT

ATTAATGAAA

CCAAAATTGA

CTTCCTGGAA

CAGACTGTTA

GGAAGTAGTC

GAAATA.AAGG

AGCAAAAGCG

TTGGCTCAGG

AGTGAGGATG

CCTTCTCAGT

GAGAATTTAT

AAGGCATCTC

TCACAGTGCA

ATTGGTTCTT

AAGGAATTGG

GAGCAAAGCA

GTCTCTGAAG

GATACCATGC

TTAGAACAGC

GCCCTTGAGG

CAGAAAAGTA

GAGGTGTCTG

CTTCTAAAT

CAGAATAGAA

CAGCTGGAAG

CTAGAGGGAA

GATCCTTCTG

3060 3120 3180 3240 3300 3360 34.20 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 AAGACAGAGC CCCAGAGTCA GCTCGTGTTG GCAACATACC ATCTTCAACC TCTGCATTGA WO 96/33271 WO 9633271PCTIUS96/05621 AAGTTCCCCA A9 ATACTGCTGG C-'j CTTCAACAGA Al AATTTATGCT CC CTGAAGAGAC T2 TGAAATATTT T( AGTCTATTAA AC TCAATGGAAG A TCAGGGGGCT AC AATGGATGGT AC GCACAGGTGT CC TCCATGCAAT TC G'TGTAGCACT C~ GCCACTICTG A 'TGAAAGTT GCAGAATCTG CCCAGAGTCC AGCTGCTGCT CATACTACTG

~ATAATGCA

~GGTCAAC

TGTACAAG

.CTCATGTT

TAGGAATT

3 AAAGPAAA k.ACCACCAA 3AAATCTGT

:AGCTGTGT

:ACCCAATT

3GGCAGATG

TACCAGTGC

ATGGAAGAAA

AAAAGAATGT

TTTGCCAGAA

GTTATGAAAA

GCGGGAGGAA

ATGCTGAATG

GGTCCAAAGC

TGCTATGGGC

GGTGCTTCTG

GTGGTTGTGC

TGTGAGGCAC

CAGGAGCTGG

GTGTGAGCAG

CCATGGTGGT

AACACCACAT

CAGATGCTGA

AATGGGTAGT

AGCATGATTT

GAGCAAGAGA

CCTTCACCAA

TGGTGAAGGA

AGCCAGATGC

CTGTGGTGAC

ACACCTACCT

GGAGAAGCCA

GTCTGGCCTG

CACTTTAACT

GTTTGTGTGT

TAGCTATTTC

TGAAGTCAGA

ATCCCAGGAC

CATGCCCACA

GCTTTCATCA

CTGGACAGAG

CCGAGAGTGG

GATACCCCAG

GAATTGACAG

ACCCCAGAAG

AATCTAATTA

GAACGGACAC

TGGGTGACCC

GGAGATGTGG

AGAAAGATCT

GATCAACTGG

TTCACCCTTG

GACAATGGCT

GTGTTGGACA

ATCCCCCACA

4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 5640 5700 5711 INFOPCMATION FOR SEQ ID NO:ll: SEQUENCE CHARACTERISTICS: LENGTH: 5707 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:ll: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG GTTTCTCAGA TAACTGGGCC

CCTGCGCTCA

TGGATTTATC

TCTTAGAGTG

ACATATTTTG

GTCCTTTATG

AACTTGTTGA

ATGCAiACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AArTGGGATC

GGAGGCCTTC

7GCTCTTCGC

TCCCATCTGT

C:AAATTTTGC

TAAGAATGAT

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAGGACA

TGATTCTTCT

ACCCTCTGCT

GTTGAAGAAG

CTGGAGTTGA

ATGCTGAAAC

ATAACCAAAA

AAAATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

AAGCAGCGGA

GAAGATACCG

'TGGGTAAAG

TACAAAATGT

TCAAGGAACC

TTCTCAACCA

GGAGCCTACA

GTGCTTTTCA

AAAATAACTC

GAAACCGTGC

GTCTCAGTGT

TACAACCTCA

TTAATAAGGC

TTCATTGGAA

CATTAATGCT

TGTCTCCACA

GAAGAAAGGG

AGAAAGTACG

GCTTGACACA

TCCTGAACAT

CAAAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

CAGAAAGAAA

ATGCAGAAAA

ALAGTGTGACC

CCTTCACAGT

AGATTTAGTC

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

.AACCTTGGAA

GTCTACATTG

AGTGTGGGAG

WO 96/33271 PCTIUS96105621 ATCAAGAATT GTTACAAATC CAAAAAAGGC TGCTTGTGAA CCAGTAATAA TGATTTGAAC ATCAGGGTAG TTCTGTTTCA GCTCATTACA GCATGAGAAC AGGCTGAATT CTGTAATAAA GGGCTGGAAG TAAGGAAACA ATCTGAATGC TGATCCCCTG CAGAGAATCC TAGAGATACT AAGTTAATGA GTGGTTTTCC GGGAGTCTGA ATCAAATGCC AATATTCTGG TTCTTCAGAG TATGTAAAAG TGAAAGAGTT TTGGGAAAAC CTATCGGAAG TAATTATAGG AGCATTTGTT AATTAAAGCG TAAAAGGAGA CAGATTTGGC AGTTCAAAAG AGAATGGTCA AGTGATGAAT CTATTCAGAA TGAGAAAAAT AAACGAAAGC TGAACCTATA ACAATTCAAA AGCACCTAAA ATGCGCTTGA ACTAGTAGTC TTGATAGTTG TTCTAGCAGT GGCACAGCAG AA.ACCTACAA GTAACAAGCC AAATGAACAG AGTTAACAAA TGCACCTGGT TTGTCAATCC TAGCCTTCCA CTAATAATGC TGAAGACCCC AAAGATCTGT AGAGAGTAGC AAAGTATCTC GTTACTGGAA GTGTGAGTCA GTGTGCAGCA ATAATAGAAA TGACACAGAA

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

TGTAATGATA

TGTGAGAGAA

GAAGATGTTC

AGAAGTGATG

AA.AGTAGCTG

AAAATAGACT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCAG

ACTCCTGAAA

ATTACTAATA

CCTAACCCAA

AGCAGCAGTA

AAGAATAGGC

AGTAGAAATC

GAAGAGATAA

CTCATGGAAG

ACAAGTAAAA

TCTTTTACTA

AGAGAAGAAA

AAAGATCTCA

AGTATTTCAT

GTTAGCACTC

TTTGARAACC

GGCTTTAAGT

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAGC

GGCGGACTCC

AAGAATGGAA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

TACTGGCCAG

CAGTAGAGAG

TCCCCAACTT

AGATAATACA

GCCTTCATCC

TGATAAATCA

GTGGTCATGA

TAGAATCACT

TAAGCAATAT

TGAGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GTAAAGAACC

GACATGACAG

AGTGTTCAAA

AAGAAGAGAA

TGTTAAGTGG

TGGTACCTGG

TGAAATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAAXAT

AGACAGAATG

AAGGAGCCAA

CAGCACAGAA

TAAGCAGAAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAAAT

TGATCCTCAT

TAATATTGAA

AAGCCATGTA

AGAGCGTCCC

TGAGGATTTT

GGGAACTAAC

GAATAAAACA

CGAAAAAGAP.

GGAACTCGAA

GTCTTCTACC

TAATTGTACT

GTACAACCAA

TGCAACTGGA

CGATACTTTC

TACCAGTGAA

ACTAGAAACA

AGAAAGGGTT

TACTGATTAT

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

AAAAAGGTAG

CTGCCATGCT

AGCATTCAGA

'CACATGATG

GAGGTAGATG

GAGGCTTTAA

GACAAAATAT

ACTGAAAATC

CTCACAAATA

ATCAAGAAAG

CAAACGGAGC

AAAGGTGATT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GAATTGCAAA

ATGCCAGTCA

GCCAAGAAGA

CCAGAGCTGA

CTTAAAGAAT

GTTAAAGTGT

TTGCAAACTG

GGCACTCAGG

CCAAATAAAT

TGTTCCAAAG

A.ACCACAGTC

780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640

I

TAGGGAAGGC AAAACAGAA CCAAGGGACT AATTCATGGT ATCCATTGGG ACATGAAGTT GGCACAGCAG AAACCTACAA CTA'1'.'U WO 96/33271 PCT/US96/05621

GGGAAACAAG

TCAAGGTTTC

AATGTGCAAC

TTGAATGTGA

AGACAGTTAA

ATGCCAAATG

ACGAAACTGG

CACCACTTTT

AAAACTTTGA

GTACAGTGAG

CAAGCAATAT

TLGGTTCCAG

ATGCTATGCT

GTAATTGTAA

ATACAGATTT

ATGCATCiCA

AAGATACTAG

TCCAGAAAGG

GTTACCGAAG

AAGAGCTTCC

CTACTAGGCA

TATCATTGAA

AGGAACATCA

GTGAATTGGA

CCAAACAAAT

TTTCAGATGA

TTCAAACTTA

CTCAGGGCTA

TAACCTGATA

GAGCCAGCCT

GCGAAATCCA

ATACCCTATA

CATAGAAATG

AAAGCGCCAG

ATTCTCTGCC

ACAAAAGGAA

TATCACTGCA

TAGTATCAAA

ACTCATTACT

TCCCATCAAG

GGAACATTCA

CACAATTAGC

TAATGAAGTA

TGATGAAAAC

TAGATTAGGG

GCATCCTGAA

CTCTCCATAT

GGTTTGTTCT

TTTTGCTGAA

AGAGCTTAGC

AGGGGCCAAG

CTGCTTCCAA

TAGCACCGTT

GAATAGCTTA

CCTTAGTGAG

AGACTTGACT

GAGGCATCAG

TGAAGAAAGA

GGTGAAGCAG

TCCTCTCAGA

1.AGCTCCAGC TCTAkCAGCT

GAACAAAGCA

AGCCAGAATC

GAAGAA.AGTG

TCATTTGCTC

CACTCTGGGT

GAAAATCAAG

GGCTTTCCTG

GGAGGCTCTA

CCAAATAAAC

TCATTTGTTA

ATGTCACCTG

CGTAATAACA

GGTTCCAGTA

ATTCAAGCAG

GTTTTGCA.AC

ATAAAAAAGC

CTGATTTCAG

GAGACACCTG

AATGACATTA

AGGAGTCCTA

AAATTAGAGT

CACTTGTTAT

GCTACCGAGT

AATGACTGCA

GAAACAAAAT

GCAAATACA.A

TCTGAAAGCC

GGAACGGGCT

CA'rCTGGGTG

GTGACATTTT

AGGAA.ATGGC

ACCCTTCCAT

CATCAGAAAA

CAGAAGGCCT

AACTTGATGC

CGTTTTCAAA

CCTTAAAGAA

GAAAGAATGA

TGGTTGGTCA

GGTTTTGTCT

ATGGACTTTT

PAACTAAATG

AAAGAGAAAT

TTAGAGAAA.A

CTAATGAAGT

AACTAGGTAG

CTGAGGTCTA

AAGAATATGA

ATAACTTAGA

ATGACCTGTT

AGGAAAGTTC

GCCCTTTCAC

CCTCAGAAGA

TTGGTAAAGT

GTCTGTCTAA

GTAACCAGGT

GTTCTGCTAG

ACACCCAGGA

AGGGAGTTGG

TGGAAGAAAA

TGAGAGTGAA

AACCACTCAG

TGAACTAGAA

CATAAGTGAC

AGCAGTATTA

TTCTGCTGAC

TCAGTATTTG

TCCAGGAAAT

ACAAAGTCCA

GTCTAATATC

GAAAGATAAG

ATCATCTCAG

ACAAAACCCA

TAAGAAAAAT

GGGAAATGAG

TGTTTTTAAA

GGGCTCCAGT

AAACAGAGGG

TAAACAAAGT

AGAAGTAGTT

ACAGCCTATG

AGATGATGGT

TGCTGTTTTT

CCATACACAT

GAACTTATCT

AAACAATATA

GAACACAGAG

AATATTGGCA

CTTGTTTTCT

TCCTTTCTTG

TCTGAGTGAC

TAAGAAGAGC

ACAAGCGTCT

CAGAGGGATA

GCTGTGTTAG

TCTTCTGCCC

ACTTCACAGA

AAGTTTGAGG

CAGAA~TACAT

GCAGAAGAGG

AAAGTCACTT

AAGCCTGTAC

CCAGTTGATA

TTCAGAGGCA

TATCGTATAC

CTGCTAGAGG

AACATTCCAA

GAAGCCAGCT

ATTAATGAAA

CCAAAATTGA

CTTCCTGGAA

CAGACTGTTA

GGAAGTAGTC

GAAATAAAGG

AGCAAAAGCG

TTGGCTCAGG

AGTGAGGATG

CCTTCTCAGT

GAGAATTTAT

AAGGCATCTC

TCACAGTGCA

ATTGGTTCTT

AAGGAATTGG

AAAGCATGGA

CTGAAGACTG

CCATGCAACA

AACAGCATGG

TTGAGGACCT

AAAGTAGTGA

TGTCTGCAGA

2700 2760 2820 2880 2940 3000 -3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 46 WO 96/33271 PTU9/52 PCT[US96/05621

TAGTTCTACC

ATCATTAGAT

CCCATCTCAA

TGGGCCACAC

TTACCTGGAA

CAGAGCCCCA

TCCCCAATTG

TGCTGGGTAT

AACAGAA.AGG

TATGCTCGTG

AGAGACTACT

ATATTTTCTA

TATTAAAGAA

TGGAAGAAAC

GGGGCTAGAA

GATGGTACAG

AGGTGTCCAC

TGCAATTGGG

AGCACTCTAC

CTACTGA

AGTAAAAATA AAGAACCAGG AGTGGAAAGG TCATCCCCTT CTAAATGCCC

GATAGGTGGT

GAGGAGCTCA

GATTTGACGG

TCTGGAATCA

GAGTCAGCTC

AA.AGTTGCAG

AATGCAATGG

GTCAACAAAA

TACAAGTTTG

CATGTTGTTA

GGAATTGCGG

AGAAAAATGC

CACCAAGGTC

ATCTGTTGCT

CTGTGTGGTG

CCAATTGTGG

CAGATGTGTG

CAGTGCCAGG

ACATGCACAG

TTAAGGTTGT

AAACATCTTA

GCCTCTTC'rC

GTGTTGGCAA

AATCTGCCCA

AAGAAAGTGT

GAATGTCCAT

CCAGAAAACA

TGAAAACAGA

GAGGAAAATG

TGAATIGAGCA

CAAAGCGAGC

ATGGGCCCTT

CTTCTGTGGT

rTGTGCAGCC

AGGCACCTGT

AGCTGGACAC

TTGCTCTGGG

TGATGTGGAG

CTTGCCAAGG

TGATGACCCT

CATACCATCT

GAGTCCAGCT

GAGCAGGGAG

GGTGGTGTCT

CCACATCACT

TGCTGAGTTT

GGTAGTTAGC

TGATTTTGAA

AAGAGAATCC

CACCAACATG

GAAGGAGCTT

AGATGCCTGG

GGTGACCCGA

CTACCTGATA

AGTCTTCAGA

GAGCAACAGC

CAAGATCTAG

GAATCTGATC

TCAACCTCTG

GCTGCTCATA

AAGCCAGAAT

GGCCTGACCC

TTAACTAATC

GTGTGTGAAC

I'ATTTCTGGG

GTCAGAGGAG

CAGGACAGAA

CCCACAGATC

TCATCATTCA

ACAGAGGACA

GAGTGGGTGT

CCCCAGATCC

GTTTCTCAGA

TTCATTGGAA

CATTAATGCT

1GTCTCCACA

GAAGAAAGGG

AGAAAGTACG

ATAGAAACTA

TGGAAGAGTC

AGGGAACCCC

CTTCTGAAGA

CATTGAAAGT

CTACTGATAC

TGACAGCTTC

CAGAAGAATT

TAATTACTGA

GGACACTGAA

TGACCCAGTC

ATGTGGTCAA

AGATCTTCAG

AACTGGAATG

CCCTTGGCAC

ATGGCTTCCA

TC-GACAGTGT

CCCACAGCCA

TAACTGGGCC

CAGAAAGAAA

ATGCAGAAAA

AAGTGTGACC

CCTTCACAGT

AGATTTAGTC

4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 5460 5520 5580 9640 5700 5707 INFORMATION FOR SEQ ID NO:12: SEQUENCE CHARACTERISTICS: LENGTH: 5712 base pairs TYPE: nucleic acid STRANDEDNESS: double TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: AGCTCGCTGA GACTTCCTGG ACCCCGCACC AGGCTGTGGG CCTGCGCTCA GGAGGCCTTC ACCCTCTGCT CTGGGTAAAG TGGATTTATC TGCTCTTCGC GTTGAAGAAG TACAAAATGT TCTTAGAGTG TCCCATCTGT CTGGAGTTGA TCAAGGAACC ACATATTTTG CAAATTTTGC ATGCTGAAAC TTCTCAACCA GTCCTTTATG TAAGAATGAT ATAACCAAAA GGAGCCTACA I U WO 96/3327 1 PCT[US96/05621

AACTTGTTGA

ATGCAAACAG

AAGTTTCTAT

AACCCGAAAA

CTGTGAGAAC

AATTGGGATC

ATCAAGAATT

CAAAAAAGGC

CCAGTAATAA

ATCAGGGTAG

GCTCATTACA

AGGCTGAATT

GGGCTGGAAG

ATCTGAATGC

CAGAGAATCC

AAGTTAATGA

GGGAGTCTGA

AATATTCTGG

TATGTAAAAG

TTGGGAAAAC

TAATTATAGG

AATTA.AAGCG

CAGATTTGGC

AGAATGGTCA

CTATTCAGAA

APLACGAAAGC

ACAATTCAAA

ATGCGCTTGA

TTGATAGTTG

GGCACAGCAG

GTAACAAGCC

AGTTAACAAA

AGAGCTATTG

CTATAATTTT

CATCCAAAGT

TCCTTCCTTG

TCTGAGGACA

TGATTCTTCT

GTTACAAATC

TGCTTGTGAA

TGATTTGAAC

TTCTGTTTCA

GCATGAGA.AC

CTGTAATAAA

TAAGGAAACA

TGATCCCCTG

TAGAGATACT

GTGGTTTTCC

ATCAAATGCC

TTCTTCAGAG

TGAAAGAGTT

CTATCGGAAG

AGCATTTGTT

TAAAAGGAGA

AGTTCAAAAG

AGTGATGAAT

TGAGAAAAAT

TGAACCTATA

AGCACCTAAA

ACTAGTAGTC

TTCTAGCAGT

AAACCTACAA

AAATGAACAG

TGCACCTGGT

AAAATCATTT

GCAAAAAAGG

ATGGGCTACA

CAGGAAACCA

AAGCAGCGGA

GAAGATACCG

ACCCCTCAAG

TTTTCTGAGA

ACCACTGAGA

AACTTGCATG

AGCAGTTTAT

AGCAAACAGC

TGTAATGATA

TGTGAGAGAA

GAAGATGTTC

AGAAGTGATG

AAAGTAGCTG

AA-AATAGACT

CACTCCAAAT

AAGGCAAGCC

ACTGAGCCAC

CCTACATCAG

ACTCCTGAAA

ATTACTAATA

CCTAACCCAA

AGCAGCAGTA

AAGAATAGGC

AGTAGAAATC

GAAGAGATAA

CTCATGGAAG

ACAAGTAAAA

TCTTTTACTA

GTGCTTTTCA

AAAANTAACTC

GAAACCGTGC

GTCTCAGTGT

TACAACCTCA

TTAATAAGGC

GAACCAGGGA

CGGATGTAAC

AGCGTGCAGC

TGGAGCCATG

TACTCACTAA

CTGGCTTAGC

GGCGGACTCC

AAGAATGGAA

CTTGGATAAC

AACTGTTAGG

ATGTATTGGA

TACTGGCCAG

CAGTAGAGAG

TCCCCAACTT

AGATAATACA

GCCTTCATCC

TGATAAATCA

GTGGTCATGA

TAGAATCACT

TAAGCAATAT

TGAGGAGGAA

TAAGCCCACC

AGAAAAAAAA

GTAAAGAACC

GACATGACAG

AGTGTTCAAA

GCTTGACACA

TCCTGAACAT

CAAAAGACTT

CCAACTCTCT

AAAGACGTCT

AACTTATTGC

TGA.AATCAGT

AAATACTGAA

TGAGAGGCAT

TGGCACAAAT

AGACAGA.ATG

ALAGGAGCCAA

CAGCACAGAA

TAAGCAGAAA

ACTAAATAGC

TTCTGATGAC

CGTTCTAAAT

TGATCCTCAT

TAATATTGAA

AAGCCATGTA

AGAGCGTCCC

TGAGGATTTT

GGGAACTAAC

GAATAAAACA

CGAAAAAGAA

GGAACTCGAA

GTCTTCTACC

TAATTGTACT

GTACAACCAA

TGCAACTGGA

CGATACTTTC

TACCAGTGAA

GGTTTGGAGT

CTAAAAGATG

CTACAGAGTG

AACCTTGGAA

GTCTACATTG

AGTGTGGGAG

TTGGATTCTG

CATCATCAAC

CCAGAAAAGT

ACTCATGCCA

AATGTAGAAA

CATAACAGAT

AAAA.AGGTAG

CTGCCATGCC

AGCATTC:AGA

TCACATGATG

GAGGTAGATG

GAGGCTTTAA

GACAAAATAT

ACTGAAA.ATC

CTCACAAATA

ATCAAGAAAG

CAAACGGAGC

AAAGGTGATT

TCTGCTTTCA

TTAAATATCC

AGGCATATTC

GAATTGCAAA

ATGCCAGTCA

GCCAAGAAGA

CCAGAGCTGA

CTTAAAGAAT

420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 WO 96/33271 PTU9/52 PCTfUS96/05621 TTGTCAATCC TAGCCTTCCA AGAGAAGAAA AAGAAGAGAA ACTAGAAACA GTTAAAGTGT

CTAATAATGC

AAAGATCTGT

AAAGTATCTC

GTGTGAGTCA

ATAATAGAAA

GGGAAACAAG

TCAAGGTTTC

AATGTGCAAC

TTGAATGTGA

AGACAGTTAA

ATGCCAAAT(;

ACGAAACT(aG

CACCACTTTT

AAAACTTTGA

GTACAGTGAG

CAAGCAATAT

TAGGTTCCAG

ATGCTATGCT

GTAATTGTAA

ATACAGATTT

ATGCATCTCA

AAGATACTAG

TCCAGAAAGG

GTTACCGAAG

AAGAGCTTCC

CTACTAGGCA

TATCATTGAA

AGGAACATCA

TGAAGACCCC

AGAGAGTAGC

GTTACTGGAA

GTGTGCAGCA

TGACACAGAA

CATAGAAATG

AAAGCGCCAG

ATTC'rCTGCC

ACAAAAGGAA

TATCACTGCA

TAGTATCAAA

ACTCATTACT

TCCCATCAAG

GGAACATTCA

CACAATTAGC

TA.ATGAAGTA

TGATGAAAAC

TAGATTAGGG

GCATCCTGAA

CTCTCCATAT

GGTTTGTTCT

TTTTGCTGAA

AGAGCTTAGC

AGGGGCCAAG

CTGCTTCCAA

TAGCACCGTT

GAATAGCTTA

CCTTAGTGAG

AAAGATCTCA

AGTATTTCAT

GTTAGCACTC

TTTGAAAACC

GGCTTTAAGT

GAAGAAAGTG

TCATTTGCTC

CACTCTGGGT

GAAAATCAAG

GGCTTTCCTG

GGAGGCTCTA

CCAAATAAAC

TCATTTGTTA

ATGTCACCTG

CGTAATAACA

GGTTCCAGTA

ATTCAAGCAG

GTTTTGCAAC

ATAAAAAAGC

CTGATTTCAG

GAGACACCTG

AATGACATTA

AGGAGTCCTA

AAATTAGAGT

CACTTGTTAT

GCTACCGAGT

AATGACTGCA

GAAACAAAAT

GCAAATACAA

TCTGAAAGCC

GGAACGGGCT

TGTTAAGTGG

TGGTACCTGG

TAGGGAAGGC

CCAAGGGACT.

ATCCATTGGG

AACTTGATGC

CGTTTTCAAA

CCTTAAAGAA

GAAAGAATGA

TGGTTGGTCA

GGTTTTGTCT

ATGGACTTTT

AAACTAAATG

AAAGAGAAAT

TTAGAGAAAA

CTAATGAAGT

AACTAGGTAG

CTGAGGTCTA

AAGAATATGA

ATAACTTAGA

ATGACCTGTT

AGGAAAGTTC

GCCCTTTCAC

CCTCAGAAGA

TTGGTAAAGT

GTCTGTCTAA

GTAACCAGGT

GTTCTGCTAG

ACACCCAGGA

AGGGAGTTGG

TGGAAGAAAA

AGAAAGGGTT

TACTGATTAT

AAAAACAGAA

AATTCATGGT

ACATGAAGTT

TCAGTATTTG

TCCAGGAAAT

ACAAAGTCCA

GTCTAATATC

GAAAGATAAG

ATCATCTCAG

ACAAAACCCA

TAAGAAAAAT

GGGAAATGAG

TGTTTTTA-L-k

GGGCTCCAGT

AAACAGAGGG

TAAACAAAGT

AGAAGTAGTT

ACAGCCTATG

AGATGATGGT

TGCTGTTTTT

CCATACACAi*

GAACTTATCT

AAACAATATA

GAACACAGAG

AATATTGGCA

CTTGTTTTCT

TCCTTTCTTG

TCTGAGTGAC

TAATCAAGAA

TTGCAA.ACTG

GGCACTCAGG

CCAAATAAAT

TGTTCCAAAG

AACCACAGTC

CAGAATACAT

GCAGAAGAGG

AAAGTCACTT

AAGCCTGTAC

CCAGTTGATA

TTCAGAGGCA

TATCGTATAC

CTGCTAGAGG

AACATTCCAA

GAAGCCAGCT

ATTAATGAAA

CCAAAATTGA

CTTCCTGGAA

CAGACTGTTA

GGAAGTAGTC

GAAATAAAGG

AGCAAAAGCG

TTGGCTCAGG

AGTGAGGATG

CCTTCTCAGT

GAGAATTTAT

AAGGCATCTC

TCACAGTGCA

ATTGGTTCTT

AAGGAATTGG

GAGCAAAGCA

2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 GTGAATTGGA AGACTTGACT CCAAACAAAT GAGGCATCAG TTTCAGATGA TGAAGAAAGA

L

WO 96/33271 PCTIUS96/05621

TGGATTCAAA

ACTGCTCAGG

AACATAACCT

ATGGGAGCCA

ACCTGCGAAA

GTGAATACCC

CAGATAGTTC

GCCCATCATT

ACTACCCATC

AGTCTGGGCC

CCCCTTACCT

AAGACAGAGC

AAGTTCCCCA

ATACTGCTGG

CTTCAACAGA

AATTTATGCT

CTGAAGAGAC

TGAAATATTT

AGTCTATTAA

TCAATGGAAG

TCAGGGGGCT

A.ATGGATGGT

GCACAGGTGT

TCCATGCAAT

GTGTAGCACT

CTTAGGTGAA CCAGCATCTG GGTGTGAGAG TGAAACAAGC GTCTCTGAAG

GCTATCCTCT

GATAAAGCTC

GCCTTCTAAC

TCCArjAZACAA

TATAAGCCAG

TACCAGTAAA

AGATGATAGG

TCAAGAGGAG

ACACGATTTG

GGAATCTGGA

CCCAGAGTCA

ATTGAAAGTT

GTATAATGCA

AAGGGTCAAC

CGTGTACAAG

TACTCATGTT

TCTAGGAATT

AGAAAGAAAA

AAACCACCAA

AGAAATCTGT

ACAGCTGTGT

CCACCCAATT

TGGGCAGATG

CTACCAGTGC

CAGAGTGACA

CAGCAGGAAA

AGCTACCCTT

AGCACATCAG

AATCCAGAAG

AATAAAGAAC

TGGTACATGC

CTCATTAAGG

ACGGAA.ACAT

ATCAGCCTCT

GCTCGTGTTG

GCAGAATCTG

ATGGAAGAAA

AAAAGAATGT

TTTGCCAGAA

GTTATGAAAA

GCGGGAGGAA

ATGCTGALATG

GGTCCAAAGC

TGCTATGGGC

GGTGCTTCTG

GTGGTTGTGC

TGTGAGGCAC

CAGGAGCTGG

TTTTAACCAC

TGGCTGAACT

CCATCATAAG

AAAAAGCAGT

GCCTTTCTGC

CAGGAGTGGA

ACAGTTGCTC

TTGTTGATGT

CTTACTTGCC

TCTCTGATGR

GCAACATACC

CCCAGAGTCC

GTGTGAGCAG

CCATGGTGGT

AACACCACAT

CAGATGCTGA

AATGGGTAGT

AGCATGATTT

GAGCAAGAGA

CCTTCACCAA

TGGTGAAGGA

AGCCAGATGC

CTGTGGTGAC

ACACCTAACC

TCAGCAGAGG

AGAAGCTGTG

TGACTCTTCT

ATTAACTTCA

TGACAAGTTT

AAGGTCATCC

TGGGAGTCTT

GGAGGAGCAA

AAGGCAAGAT

CCCTGAATCT

ATCTTCAACC

AGCTGCTGCT

GGAGA6AGCCA

GTCTGGCCTG

CACTTTAACT

GTTTGTGTGT

TAGCTATTTC

TGAAGTCAGA

ATCCCAGGAC

CATGCCCACA

GCTTTCATCA

CTGGACAGAG

CCGAGAGTGG

TGATACCCCA

',ATACCATGC

TTAGAACAGC

GCCCTTGAGG

CAGAAA.AGTA

GAGGTGTCTG

CCTTCTAAAT

CAGAATAGAA

CAGCTGGAAG

CTAGAGGGAA

GATCCTTCTG

TCTGCATTGA

CATACTACTG

GAATTGACAG

ACCCCAGA6AG

AATCTAATTA

GAACGGACA C

TGGGTGACCC

GGAGATGTGG

AGAAAGATCT

GATCAACTGG

'IiTCACCCTTG

GACAATGGCT

GTGTTGGACA

GATCCCCCAC

4260 4320 4380 4 44 0 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 51.60 5220 5280 5340 5400 5460 5520 5580 5640 5700 5712 AGCCACTACT GA INFORMATION FOR SEQ ID NO:13: Wi SEQUENCE CHARACTERISTICS: LENGTH: 26 amino acids TYPE: amino acid STRANDEDNESS., single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: 1 i WO 96/33271 PCTIUS96/05621 Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gin Asn Val Ile Asn 1 5 10 Ala Met Gin Lys Ile Leu Glu Cys Pro Ile INFORMATION FOR SEQ ID NO:14: SEQUENCE CHARACTERISTICS: LENGTH: 38 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gin Asn Val Ile Asn 1 5 10 Ala Met Gin Lys Ile Leu Glu Cys Pro Ile Cys Leu Glu Leu Ile Lys 25 Glu Pro Val Ser Thr Val INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 63 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val 1 5 10 Ala Met Gln Lys Ile Leu Glu Cys Pro Ile Cys 25 Glu Pro Val Ser Thr Lys Cys Asp His Ile Phe 40 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Ser 55 INFORMATION FOR SEQ ID NO:16: SEQUENCE CHARACTERISTICS: LENGTH: 1863 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Gin Asn Val Ile Asn Leu Glu Leu Ile Lys Cys Lys Phe Cys Met Gin Cys Pro Leu

I

low

I

WO 96/33271 Met Asp 1 Ala Met Glu Pro Leu Lys Lys Asn Gin Leu Thr Gly Asn Ser Gly Tyr 130 Pro Ser 145 Thr Val Ser Val Lys Ala Pro Gin 210 Ala Cys 225 Pi:o Ser His Pro Pro Cys Ser Leu 290 Cys Asn 305 Trp, Ala Leu Gin Val Leu Asp Val1 Leu Pro 115 Arg Leu Arg Tyr Thr 195 Gly Giu Asn Giu Gly 275 Leu Lys G ly PCTIUS96/05621 I Ile Asn .i Ile Lys 2 Cys Met Leu Cys ;Phe Ser i Leu Asp s Giu Asn i Ser Met )Giu Asn a Leu Gly 160 i Lys Thr 175 Val Asn SIle Thr 3 Lys Ala 3 His Gin 240 i Giu Arg 255 5 Val Giu i Asn Ser i. Giu Phe 3 Asn Arg 320 Ser Thr 335 52 .3

L

WO 96/33271 Giu Lys Lys Val Asp Leu Asn Ale 340 PCTIUS96/05621 Arg Lys Giu 350 Asp Pro Leu Cys Giu 345 Trp Asp Trp 385 Gly Asn Ala Ser 'y r 465 Leu Pro His Pro Val.

545 Ser Giu Asn Asn Leu 625 Ile Gin Gin Trp Arg Giu Asp 420 Pro Val Lys Gly Asn 500 Asp Ile Ile Asn Phe 580 I.Su Arg Ser Cys Val.

660 Cys 360 Asn Leu Lys

C,Y

Leu 440 Ile Pro Thr Arg Lys 520 Thr Gly Pro Ala Ile 600 Ser Ser Giu Arg Giu Giu Asp 400 Leu Leu His Thr Asn 480 Arg Leu Thr Gin Asp 560 Lys Ser Lys Giu Gin 640 Asi.

Lys WO 96/33271 PCTUS96/05621 Glu Pro Ala Thr Gly Ala Lys Lys Ser Asn Lys Pro Asn Glu Gln Thr 675 680 685 Ser Lys Arg His Asp Ser Asp Thr Phe Pro Glu Leu Lys Leu Thr Asn 690 695 700 Ala Pr,- Gly Ser Phe Thr Lys Cys Ser Asn Thr Ser Glu Lou Lys Glu 705 710 715 720 Phe Val Asn Pro Ser Leu Pro Arg Glu Glu Lys Glu Glu Lys Leu Glu 725 730 735 Thr Val uys Val Ser Asn Asn Ala Glu Asp Pro Lys Asp Leu Met Leu- 740 745 750 Ser Gly Glu Arg Val Leu Gln Thr Glu Arg Ser Val Glu Ser Ser Ser 755 760 765 Ile Ser Leu Val Pro Gly Thr Asp Tyr Gly Thr Gin Glu Ser Ile Ser 770 775 780 Leu Leu Glu Val Ser Thr Leu Gly Lys Ala Lys Thr Glu Pro Asn Lys 785 790 795 800 Cys Val Ser Gin Cys Ala Ala Phe Glu Asn Pro Lys Gly Leu Ile His 805 810 815 Gly Cys Ser Lys Asp Asn Arg Asn Asp Thr Glu Gly Phe Lys Tyr Pro 820 825 830 Leu Gly His Glu Val Asn His Ser Arg Glu Thr Ser Ile Glu Met Glu 835 840 845 Glu Ser Glu Leu Asp Ala Gin Tyr Leu Gin Asn Thr Phe Lys Val Ser 850 855 860 Lys Arg Gin Ser Phe Ala Pro Phe Ser Asn Pro Gly Asn Ala Glu Glu 865 870 875 880 Glu Cys Ala Thr Phe Ser Ala His Ser Gly Ser Leu Lys Lys Gln Ser 885 890 895 Pro Lys Val Thr Phe Glu Cys Glu Gin Lys Glu Glu Asn Gin Gly Lys 900 905 910 Asn Glu Ser Asn Ile Lys Pro Val Gln Thr Val Asn Ile Thr Ala Gly 935 920 925 Phe Pro Val Val Gly Gin Lys Asp Lys Pro Val Asp Asn Ala Lys Cys 930 935 940 Ser Ile Lys Gly Gly Ser Arg Phe Cys Leu Ser Ser Gln Phe Arg Gly 945 950 955 960 Asn Glu Thr Gly Leu Ile Thr Pro Asn Lys His Gly Leu Leu Gin Asn 965 970 975 Pro Tyr Arg Ile Pro Pro Leu Phe Pro Ile Lys Ser Phe Val Lys Thr 980 985 990 Lys Cys Lys Lys Asn Leu Leu Glu Glu Asn Phe Glu Glu His Ser Met 995 1000 1005 54 1b 41 WO 9633271PCTJUS96/05621 Ser Pro Giu Arg Giu Met Gly Asn Giu TAsn Ile Pro Ser Thr Val Ser 1010 1015 1020 Thr Ile Ser Arg Asn Asn Ile Arg Giu Asn Val Phe Lys Giu Ala Ser 1025 1030 1035 1040 Ser Ser Asn Ile Asn Giu Val Gly Ser Ser Thr Asn Giu Val Gly Ser 1045 1050 1055 Ser Ile Asn Giu Ile Gly Ser Ser Asp Giu Asr Ile Gin Ala Giu Leu 1060 1065 1070 G'y Arg Asn Arg Gly Pro Lys Leu Asn Ala Met Leu Arg Leu Gly Val 1075 1080 1085 Leu Gin Pro Glu Val Tyr Lys Gin Ser Leu Pro Gly Ser Asn Cys Lys 1090 1095 1100 H.I.s Pro Giu Ile Lys Lys Gin Giu Tyr Giu Giu Vai Val Gin Thr Val 105 1110 1115 1120 Asn Thr Asp Phe Ser Pro Tyr Leu Ile Ser Asp Asn Leu Giu Gin Pro 1125 1130 1135 Met Gly Ser Ser His Ala Ser Gin Val Cys Ser Giu Thr Pro Asp Asp 1.140 1145 1150 Leu Leu Asp Asp Gly Giu Ile Lys Giu Asp Thr Ser Phe Ala Giu Asn 301151101E Asp Ile Lys Giu Ser Ser Ala Val Phe Ser Lys Ser Val Gin Lys Gly 1170 1175 1180 Giu Leu Ser Arg Ser Pro Ser Pro Phe Thr His Thr His Leu Ala Gin 1185 1190 1195 1200 Gly Tyr Arg Arg Gly Ala Lys Lys Leu Giu Ser Ser Giu Giu Asn Leu 1205 1210 1215 Ser Ser Glu Asp Giu Giu Leu Pro Cys Phe Gin His Leu Leu Phe Gly 1220 1225 1230 Lys Val Asn Asn Ile Pro Ser Gin Ser Thr Arg His Ser Thr Val Ala 451235 1240 1245 Thr Giu Cys Leu Ser Lys Asn Thr Giu Giu Asn Leu Leu Ser Leu Lys 1250 1255 1260 Asn Ser Leu Asn Asp Cys Ser Asn Gin Vai Ile Leu Ala Lys Ala Ser 1265 1270 1275 1280 Gin Giu His His Leu Ser Giu Glu Thr Lys Cys Ser Ala Ser Leu Phe 1285 1290 1295 Ser Ser Gin Cys Ser Glu Leu Giu Asp Leu Thr Ala Asn Thr Asn Thr 1300 1305 1310 Asp Pro e Leu Ile Gly Ser Ser Lys Gin Met Arg His Gin Ser 601315 1320 1325 Giu Ser Gin Gly Val Gly Leu Ser Asp Lys Giu Leu Val Ser Asp Asp 1330 1335 1340 42 WO 96/33271 PCTIUS9605621 Glu Glu Arg Gly Thr Gly Leu Glu Glu Asn Asn Gin Glu Glu Gin Ser 1345 1350 1355 1360 Met Asp Ser Asn Leu Gly Glu Ala Ala Ser Gly Cys Glu Ser Glu Thr 1365 1370 1375 Ser Val Ser Glu Asp Cys Ser Gly Leu Ser Ser Gin Ser Asp Ile Leu 1380 1385 1390 Thr Thr Gin Gin Arg Asp Thr Met Gin His Asn Leu Ile Lys Leu Gin 1395 1400 1405 Gin Glu Met Ala Glu Leu Glu Ala Val Leu Glu Gin His Gly Ser Gin 1410 1415 1420 Pro Ser Asn Ser Tyr Pro Ser Ile Ile Ser Asp Ser Ser Ala Leu Glu 1425 1430 1435 1440 Asp Leu Arg Asn Pro Glu Gin Ser Thr Ser Glu Lys Ala Val Leu Thr 1445 1450 1455 Ser Gin Lys Ser Ser Glu Tyr Pro Ile Ser Gin Asn Pro Glu Gly Leu 1460 1465 1470 Ser Ala Asp Lys Phe Glu Val Ser Ala Asp Ser Ser Thr Ser Lys Asn 1475 1480 1485 Lys Glu Pro Gly Val Glu Arg Ser Ser Pro Ser Lys Cys Pro Ser Leu 1490 1495 1500 Asp Asp Arg Trp Tyr Met His Ser Cys Ser Gly Ser Leu Gin Asn Arg 1505 1510 1515 1520 Asn Tyr Pro Ser Gin Glu Glu Leu Ile Lys Val Val Asp Val Glu Glu 1525 1530 1535 Gin Gin Leu Glu Glu Ser Gly Pro His Asp Leu Thr Glu Thr Ser Tyr 1540 1545 1550 Leu Px, Arg Gin Asp Leu Glu Gly Thr Pro Tyr Leu Glu Ser Gly Ile 1555 1560 1565 Ser Leu Phe Ser Asp Asp Pro Glu Ser Asp Pro Ser Glu Asp Arg Ala 1570 1575 1580 Pro Glu Ser Ala Arg Val Gly Asn Ile Pro Ser Ser Thr Ser Ala Leu 1585 1590 1595 1600 Lys Val Pro Gin Leu Lys Val Ala Glu Ser Ala Gin Cer Pro Ala Ala 1605 1610 1615 Ala His Thr Thr Asp Thr Ala Gly Tyr Asn Ala Met Glu Glu Ser Val 1620 1625 1630 Ser Arg Glu Lys Pro Glu Leu Thr Ala Ser Thr Glu Arg Val Asn Lys 1635 1640 1645 Arg Met Ser Met Val Val Ser Gly Leu Thr Pro Glu Glu Phe Met Leu 1650 1655 1660 Val Tyr Lys Phe Ala Arg Lys His His Ile Thr Leu Thr Asn Leu Ile 1665 1670 1675 1680 Ir U-Y -YI 11~- CI bl r Y L-~I WO 96133271 PCTUS96/05621 Thr Giu Giu Thr Thr His Val Val Met Lys Thr Asp Ala Giu Phe Val 1685 1690 1695 Cys Giu Arg Thr Leu Lys Tyr Phe Leu Gly Ile Ala Gly Gly Lys Trp 1700 1705 1710 Val Val Ser Tyr Phe Trp Val Thr Gin Ser Ile Lys Giu Arg Lys Met 1715 1720 1725 Leu Asn Giu His Asp Phe Giu Val Arg Gly Asp Val Vai Asn Gly Arg 1730 1735 1740 Asn His Gln Gly Pro Lys Arg Ala Arg Giu Ser Gin Asp Arg Lys Ile 1745 1750 1755 1760 Phe Arg Giy Leu Glu Ile Cys Cys Tyr Gly Pro Phe Thr Asn Met Pro 1765 1770 1775 Thr Asp Gin Leu Giu Trp Met Val Gin Leu Cys Gly Ala Ser Val Val 1780 1785 1790 Lys Giu Leu Ser Ser Phe Thr Leu. Gly Thr Giy Val His Pro Ile Val 1795 1800 1805 Vai Val Gin Pro Asp Ala Trp Thr GJu Asp Asn Gly Phe His Ala Ile 1810 1815 1820 Gly Gin Met Cys Glu Ala Pro Val Val. Thr Arg Giu Trp Val Leu Asp 1825 1830 1835 1840 Ser Val Ala Leu Tyr Gin Cys Gin Giu Leu Asp Thr Tyr Leu Ile Pro 1845 1850 1855 Gin Ile Pro His Ser His Tyr 1860 INFORMATION FOR SEQ ID NO:17: SEQUENCE CHARACTERISTICS: LENGTH: 80 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: Met Asp Leu Ser Ala Leu Arg Val Giu Giu Val Gln Asn Val Ile Asn 1 5 10 15 Ala Met Gin Lys Ile Leu Giu Cys Pro Ile Cys Leu Giu Leu Ile Lys 25 30 Glu Pro Val Ser Thr Lys Cys Asp His Ile Phe Cys Lys Phe Cys Met 40 45 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Ser Gin Cys Pro Leu Cys 55 60 Lys Asn Asp Ile Thr Lys Ser Val Leu Lys Arg Leu Ile Ile Thr Cys 70 75 80 WO 961332,

INFOR

SE

(ii) M (xi) S Met A 1 Ala M Glu P Leu L Lys A Gin Thr Asn Gly Pro 145 Thr Ser Lys Pro Ala 225 Pro His PCTfUS9/O5621 WO 961332i 1 INFORMATION FOR SEQ ID NO:18: SEQUENCE CHARACTERISTICS: LENGTH: 312 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: PCT[US96105621 Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gln Asn Val Ile Asni Ala Glu Leu Lys Gln Thr Asn Gly Pro 145 Thr Ser Lys Pro Ala 225 Pro His Leu Lys Gln Lys 70 Leu Ala Ala Thr 150 Arg Leu Ser Asp Glu 230 Leu Gln Lys Met Cys Ser Asp Asn Met Asn Gly 160 Thr Asn Thr Al a Gln 240 Arg Glu -jo WO 96/33271 PCT/US96/05621 Pro Cys Gly Thr Asn Thr His Ala Ser Ser Leu Gin His Glu Asn Ser 275 280 285 Ser Leu Leu Leu Thr Lys Asp Arg Met Asn Val Glu Lys Ala Glu Phe 290 295 300 Cys Asn Lys Ser Lys Arg Leu Ala 305 310 INFORMATION FOR SEQ ID NO:19: SEQUENCE CHARACTERISTICS: LENGTH: 765 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gin Asn Val Ile Asn 1 5 10 Ala Met Gin Lys Ile Leu Glu Cys Pro Ile Cys Leu Glu Leu Ile Lys 25 Glu Pro Val Ser Thr Lys Cys Asp His Ile Phe Cys Lys Phe Cys Met 40 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Ser Gin Cys Pro Leu Cys 55 Lys Asn Asp Ile Thr Lys Arg Ser Leu Gin Glu Ser Thr Arg Phe Ser 65 70 75 Gin Leu Val Glu Glu Leu Leu Lys Ile Ile Cys Ala Phe Gin Leu Asp 90 Thr Gly Leu Glu Tyr Ala Asn Ser Tyr Asn Phe Ala Lys Lys Glu Asn 100 105 110 Asn Ser Pro Glu His Leu Lys Asp Glu Val Ser Ile Ile Gin Ser Met 115 120 125 Gly Tyr Arg Asn Arg Ala Lys Arg Leu Leu Gin Ser Glu Pro Glu Asn I 130 135 140 Pro Ser Leu Gin Glu Thr Ser Leu Ser Val Gin Leu Ser Asn Leu Gly 145 150 155 160 Thr Val Arg Thr Leu Arg Thr Lys Gin Arg Ile Gin Pro Gin Lys Thr 165 170 175 Ser Val Tyr Ile Glu Leu Gly Ser Asp Ser Ser Glu Asp Thr Val Asn 180 185 190 Lys Ala Thr Tyr Cys Ser Val Gly Asp Gin Glu Leu Leu Gin Ile Thr 195 200 205 Pro Gin Gly Thr Arg Asp Glu Ile Ser Leu Asp Ser Ala Lys Lys Ala 210 215 220 59 a 46 WO 96/33271 PCT/US96/05621 Ala Cys Giu Phe Ser Glu Thr Asp Val. Thr Asn Thr Giu His His Gin 225 230 235 240 4G 55 Asn Giu Gly 275 Leu Lys G ly Lys Lys 355 Pro Ser Ser Val Asp 435 Ser Lys Ile Thr Giu 515 Met Asn Asn Lys 260 Thr Leu Ser Ser Val 340 Gin Trp Arg Giu Asp 420 Pro Val.

Lys Gly Asn 500 Asp Ile Ile Leu Gin Thr Lys Gin 310 Giu Leu Leu Thr Asp 390 Asn Tyr Giu Ser Ser 470 Phe Leu Ile Gin Asn 550 Asn Giy His Asp 295 Pro Thr Asn Pro Leu 375 Glu Ala Ser Ala Asn 455 Leu Val Lys Lys Giy 535 Ser Thr Ser 265 Ser Me t Leu Asn Asp 345 Ser Ser Leu Vai Ser 425 Ile Glu Asn Giu Lys 505 Ala Asn His Giu 250 Vai Ser Asn Aia Asp 330 Pro Giu Ser Gly Ala 410 Ser Cys Asp Leu Pro 490 Arg Asp Gin Arg Asn Gin Giu 300 Ser Arg Cys Pro 380 Asp Val Lys Ser Ile 460 His Ile Pro Aia Glu 540 Aia Leu His 285 Lys Gin Thr Giu Arg

CS-

Lys Asp Leu Ile Giu 445 Ph Vai Ile Thr Vai 525 Gin Aia His 270 Giu Ala His Pro Arg 350 Asp Val Ser Asp Asp 430 Arg Giy Thr Gin Ser 510 Gin Asn Arg Giu Ser Phe Arg 320 Thr Giu Giu Giu Asp 400 Leu Leu His Thr Asn 480 Arg Leu Thr Gin

'I

Giu Asn Lys Thr Lys Giy 555 WO 96/33271 Ser Ile Giu Ser Asn Met Asn Arg 610 Leu Val 625 Ile Asp Gin Met Giu Pro Ser Lys 690 Ala Pro 705 Phe Vai Thr Val Giu Ser Lys Ile 620 Cys Lys Leu Pro Leu 700 Ser Giu Lys Vali PCT[US96/05621 Leu Giu Lys 575 5cr Ile Ser 590 Pro Lys Lys Ala Leu Giu Giu Leu Gin 640 Lys Tyr Asn 655 Giu Ciy Lys 670 Giu Gin Thr Leu Thr Asn Leu Lys Giu 720 Lys Leu Glu 735 Leu Met Leu 750 Ser Gly Arg Vai Leu Gin Thr 760 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: (ii) (xi) Met 1 Ala Glu Leu LENGTH: 900 amino acids TYPE: amino acid STRAN~DEDNESS: single TOPOLOGY: linear MOLECULE TYPE: protein SEQUENCE DESCRIPTION: SEQ ID Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gin Asn Vai Ile As,, 5 10 Met Gin Lys Ile Leu Giu Cys Pro Ile Cys Leu Glu Leu Ile Lys 25 Pro Val Ser Thr Lys Cys Asp His Ile Phe Cys Lys Phe Cys Met 40 Lys Leu Leu Asn Gin Lys Lys Gly Pro 5cr Gin Cys Pro Leu Cys 55 61 AGTTAACAAA TGCACCTGGT TCTTTTACTA AGTGTTCAAA TACCAUTUIA~ i '~TTAAG TTTCAGATGA T WO 96/33271 Lys Asn Asp Ile PCTIUS96/05621 Arg Ser Leu Gin Giu Ser Thr Arg Phe Ser WO 96/33271 Gly C- Thr Lys Gin Thr Asn Giy Pro 145 Thr Ser LysJ Pro( Aia 225 Pro HisI ProC Ser1 CysP 305 Trp TrpA 'spV TrpP 385 Leu Giy Ser Tyr 130 Ser Vai Vai Aia Gin 210 Cys Ser Pro Cys Leu 290 Asn Aia Lys Asn Vai 370 Phe Vai Leu Pro 115 Arg Leu Arg Tyr Thr 195 Giy Giu Asn Giu Giy 275 Leu Lys Giy Lys Lys 355 Pro SerJ Giu Giu 100 Giu Asn Gin Thr Ile 180 Tyr Thr Phe Asn Lys 260 Thr Leu Ser Ser Vai 340 Gin Trp Arg Giu Leu Tyr Aia His Leu Arg Aia Giu Thr 150 Leu Arg 165 Giu Leu Cys Ser Arg Asp Ser Giu 230 Asp Leu 245 Tyr Gin Asn Thr Thr Lys Lys Gin 310 Lys Giu 325 Asp Leu Lys LeuI Ile ThrI Ser Asp 390 Leu Asn Lys Lys i3 5 Ser Thr Giy Vai Giu 215 Thr Asn Giy His Asp 295 Pro Thr Asn ProI Leu 375 GiuI Lys Ser Asp 120 Arg Leu Lys Ser Giy 200 Ile Asp Thr Ser Aia 280 ArgI GiyI Cysj AiaJ Cys 360 Asn LeuI Ile Tyr 105 Giu Leu I S erI Asp 185 Asp SerI ValI ThrC 265 MetP LeuP AsnP AspF 345 Ser G SerS LeuG Ile 90 Asn Vai Leu Vai Arg 170 Ser Gin Leu Thr Giu 250 Vai Ser Asn Aia Asp 330 ProI Giu Ser !Cys LPhe Ser Gin Gin Ilie Ser GiuI Asp Asn1 235 LysI SerI LeuC Vai Arg S 315 ArgP LeuC AsnF IleG Aia Aia Ile S er i4 0 Leu Gin Giu Leu Ser 220 Thr Arg Asn Gin Giu 300 Ser Arg Cys Pro GinI LPhe LLys Ile i2 Ser Pro Asp Leu 205 AiaI GiuI Aiaj LeuI His 285 LysI GinI ThrI Giu ArgP 365 Lys N Gin Lys Gin Pro Asn Gin Thr 190 Gin Lys His Aia His 270 Giu Aia His Pro Arg 350 Asp Val LLeu Giu Ser Giu Leu Lys 175 Vai Ile Lys His Giu 255 Vai Asn Giu Asn Ser 335 Lys Thr Asn IAsp iAsn 7Met Asn- Giy 160 Thr Asn Thr Aia Gin 240 Arg Giu Ser Phe Arg 320 Thr Giu Giu Giu I110 Asn C Aia S Ser I Tyr 1 465 Leu Pro His Pro Vai 545 Ser Giu Asn Asn Leu 625 Ile Gin Giu Ser Aia 705 ~a 380 Giy Ser 395 Asp Asp Ser His

I

TTITCAGATGA TGAAGAAAGA GGAACGGGCT TGGAAGAAAA. TAATCAAGA GAGCAAAGCA 4200 j WO 96/33271 PCTJUS96/05621 Gly Glu Ser Giu Ser Asn Ala Lys Val Ala Asp Val Leu Asp Val Leu 405 410 415 Asn Ala Ser Tyr 465 Leu Pro His Pro Val 545 Ser Giu Asn Asn Leu 625 Ile Gin Giu Ser Ala 705 Phe Giu Ser Lys 450 Arg Ile Leu Pro Giu 530 Met Ile S er Met Arg 610 Val Asp Met Pro Lys 690 prQ, Val Glu His Glu Ala Ala 485 Lys Phe Asn Thr Giu 565 Lys Giu Arg Arg Ser 645 Arg Gly Asp Phe Sr 725 Gly Leu 440 Ile Pro Thr Arg Lys 520 Thr Gly Pro Ala Ile 600 Ser Ser Giu Arg Lys 680 Thr Cys Arg Asp 430 Arg Giy Thr Gin Ser 510 Gin Asn Lys Leu Ser 590 Pro Ala Giu Lys Giu 670 Giu Leu Leu Lys

I

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: WO 96/33271 Thr Val Lys Val Ser Asn Asn Ala Glu Asp 740 745 Ser Gly Glu Arg Val Leu Gln Thr Glu Arg 755 760 Ile Ser Leu Val Pro Gly Thr Asp Tyr Gly 770 775 Leu Leu Glu Val Ser Thr Leu Gly Lys Ala 785 790 Cys Val Ser Gln Cys Ala Ala Phe Glu Asn 805 810 Gly Cys Ser Lys Asp Asn Arg Asn Asp Thr 820 825 Leu Gly His Glu Val Asn His Ser Arg Glu 835 840 Glu Ser Glu Leu Asp Ala Gln Tyr Leu Gln 850 855 Lys Arg Gln Ser Phe Ala Pro Phe Ser Asn 865 870 Glu Cys Ala Thr Phe Ser Ala His Ser Gly 885 890 Lys Ser His Phe 900 INFORMATION FOR SEQ ID NO:21: SEQUENCE CHARACTERISTICS: LENGTH: 914 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: Met Asp Leu Ser Ala Leu Arg Val Glu Glu 1 5 10 Ala Met Gln Lys Ile Leu Glu Cys Pro Ile 20 Glu Pro Val Ser Thr Lys Cys Asp His Ile 35 Leu Lys Leu Leu Asn Gln Lys Lys Gly Pro Lys Asn Asp Ile Thr Lys Arg Ser Leu Gln 70 Gln Leu Val Glu Glu Leu Leu Lys Ile Ile 90 Pro Ser Thr Lys 795 Pro Glu Thr Asn Pro 875 Ser Val Cys Phe Ser Glu Cys PCT/US96/05621 Leu Met Leu 750 Ser Ser Ser Ser Ile Ser Pro Asn Lys 800 Leu Ile His 815 Lys Tyr Pro 830 Glu Met Glu Lys Val Ser Ala Glu Glu 880 Thr Lys Ser 895 ^r Ile Ile Cys Leu Phe Leu WO 96/3327 1 Thr G Asn S Gly T 1 Pro S 145 Thr V Ser V Lys A P ro G 2 Ala C 225 Pro S His P Pro C Ser L 2 Cys A 305 Trp A Giu L Trp A Asp V 3 Trp P 385 Gly G Asn G PCT/US96/05621 Lys Glu Asn 110 Gin Ser Met Pro Glu Asn Asn Leu Gly 160 Gin Lys Thr 175 Thr Val Asn 190 Gin Ile Thr Lys Lys Ala His His Gin 240 Ala Giu Arg 255 His Val Giu 270 Giu Asn Ser Ala Giu Phe His Asn Arg 320 Pro Ser Thr 335 Arg Lys Giu 350 Asp Thr Giu Val Asn Giu Ser His Asp 400 Asp Vai Leu 415 Asp Leu Leu 430 WO 96/33271 PC' )6/05621 Ala Ser Asp Pro His Giu Ala Leu Ile Cys Lys Ser Giu Arg Val His 435 440 445 Ser Lys Ser Val Glu Ser Asn Ile Giu Asp Lys Ile Phe Gly Lys Thr 4590 455 460 Tyr Arg Lys Lys Ala Ser Leu Pro Asn Leu Ser His Val Thr Glu Asn 465 470 475 480 Leu Ile Ile Gly Ala Phe Val Thr Giu Pro Gln Ile Ile Gin Giu Arg 485 490 495 Pro Leu Thi Asn Lys Leu Lys Arg Lys Arg Arg Pro Thr Ser Gly Leu- 500 505 510 His Pro Giu Asp Phe Ile Lys Lys Ala Asp Leu Ala Val Gin Lys Thr 515 520 525 Pro Glu Met Ile Asn Gin Gly Thr Asn Gin Thr Giu Gin Asn Gly Gin 530 535 540 Val Met Asn Ile Thr Asn Ser Gly His Giu Asn Lys Thr Lys Gly Asp 545 550 555 560 Ser Ile Gin Asn Giu Lys Asn Pro Asn Pro Ile Giu Ser Leu Giu Lys 565 570 575 Giu Ser Ala Phe Lys Thr Lys Ala Giu Pro Ile Ser Ser Ser Ile Ser 580 585 590 Asn Met Giu Leu Giu Leu Asn Ile K.As Asn S3er Lys Ala Pro Lys Lys 595 600 605 Asn Arg Leu Arg Arg Lys Ser Ser Thr Arg His Ile His Ala Leu Giu 610 615 620 Leu Vai Val Ser Arg Asn Leu Ser Pro Pro Asn Cys Thr Giu Leu Gin 625 630 635 640 Ile Asp Ser Cys Ser Ser Ser Glu Giu Ile Lys Lys Lys Lys Tyr Asn 645 650 655 Gin Met Pro Vai Arg His Ser Arg Asr~ Leu Gin Leu Met Giu Giy Lys 45660 665 670 Giu Pro Ala Thr Giy Ala Lys Lys Ser Asn Lys Pro Asn Giu Gin Thr 675 680 685 Ser Lys Arg His Asp Ser Asp Thr Phe Pro Giu Leu Lys Leu Thr Asn 690 695 700 Ala Pro Giy Ser Phe Thr Lys Cys Ser Asn Thr Ser Giu Leu Lys Giu 705 710 715 720 Phe Val Asn Pro Ser Leu Pro Arg Giu Giu Lys Glu Giu Lys Leu Giu 725 730 735 Thr Vai Lys Ser Asn Asn Ala Glu Asp Pro Lys Asp Leu Met Leu 60740 745 750 Ser Gly Giu Arg Vai Leu Gin Thr Glu Arg Ser Val Giu Ser Ser Ser 755 760 765 000 b/U WO 96/33271 Ile Ser Leu Val Pro Gly Thr Asp Tyr Gly 770 775 Leu Leu Glu Val Ser Thr Leu Gly Lys Ala 785 790 Cys Val Se; Gln Cys Ala Ala Phe Glu Asn 805 810 Gly Cys Ser Lys Asp Asn Arg Asn Asp Thr 820 825 Leu Gly His Glu Val Asn His Ser Arg Glu 835 840 Glu Ser Glu Leu Asp Ala Gln Tyr Leu Gin 850 855 Lys Arg Gin Ser Phe Ala Pro Phe Ser Asn 865 870 Glu Cys Ala Thr Phe Ser Ala His Ser Gly 885 890 Pro Lys Val Thr Pne Glu Cys Glu Gin Lys 900 905 Asn Glu INFORMATION FOR SEQ ID NO.22: SEQUENCE CHARACTERISTICS: LENGTH: 1202 amino acids 33 TYPE: e.mino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: Met Asp Leu Ser Ala Leu Arg Val Glu Glu 1 5 Ala Met Gin Lys Ile Leu Glu Cys Pro Ile Glu Prn Val Ser Thr Lys Cys Asp His Ile 35 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Lys Asn Asp Ile Thr Lys Arg Ser Leu Gin Gin Leu Val Glu Glu Leu Leu Lys Ile Ile Thr Gly Leu Glu Tyr Ala Asn Ser Tyr Asn 100 105 PCT/US96/05621 Ser Ile Ser Pro Asn Lys 800 Leu Ile His 815 Lys Tyr Pro 830 Glu Met Glu Lys Val Ser Ala Glu Glu 880 Lys Gin Ser 895 Gin Cly Lys 910 67 ~u

I

1000 1005 WO 96/33271 Asn Ser Pro Glu His Leu Lys 115 PCT1US96/05621 Giu Val. Ser Ile Ile Gin Ser Met 125 Arg Leu Arg Tyr Thr 195 Giy Giu Asn Giu Gly 275 Leu Lys Giy Lys Lys 355 Pro Ser Ser Vai Asp 435 Leu Val1 Arg 170 Ser Gin Leu Thr Giu 2 Val Ser Asn Ala Asp 330 Pro G iu Ser Gly Ala 410 Ser Cys Glu Pro Giu Ser Pro Asp Leu 205 Ala Glu Ala Leu His 285 Lys Gin Thr Glu Arg 365 Lys Asp Leu Ile Glu 445 Asn Gin Thr 190 Gin Lys His Aia His 270 Giu Aia His Pro Arg 350 Asp Val Ser Asp Asp 430 Arg Asn Giy 160 Thr Asn Thr Ala Gin 240 Arg Giu Ser Phe Arg 320 Thr Giu Glu Giu Asp 400 Leu Leu His 153U1335 14 1340 WO096/33,: -1 Ser I Tyr 1 465 Leu Pro I His I Pro Val 1 545 Ser Giu Asn I Asn I Leu 1 625 Ile I Gin I GiuI Ser I Ala 1 705 Phe I ThrI SerC Ile Ile 460 His Ile Pro Ala Giu 540 Lys Giu Ser Lys Ile 620 Cys Lys Leu Pro Leu 700 Ser Giu Lys Val Gin 780 PCTIUS96/05621 Giy Lys Thr Thr Giu Asn 480 Gin Giu Arg 495 Ser Giy Leu 510 Gin Lys Thr Asn Gly Gin Lys Giy Asp 560 Leu Giu Lys 575 Ser Ile Ser 590 Pro Lys Lys Aia Leu Giu Giu Leu Gin 640 Lys Tyr Asn 655 Giu Giy Lys 670 Giu Gin Thr Leu Thr Asn Leu Lys Giu 720 Lys Leu Giu 735 Leu Met Leu 750 Ser Ser Ser Ser Ile Ser 1 (59 1665 1670 1675 160 56 WO 96/33271 PCTUS96/05621 Leu Leu Glu Val Ser Thr Leu Gly Lys Ala Lys Thr Glu Pro Asn Lys 785 790 795 800 Cys Val Ser Gin Cys Ala Ala Phe Glu Asn Pro Lys Gly Leu Ile His 805 810 815 Gly Cys Ser Lys Asp Asn Arg Asn Asp Thr Glu Gly Phe Lys Tyr Pro 820 825 Leu Gly His Glu Val Asn His Ser Arg Glu Thr Ser Ile Glu Met Glu 835 840 845 Glu Ser Glu Leu Asp Ala Gin Tyr Leu Gin Asn Thr Phe Lys Val Ser- 850 855 860 Lys Arg Gin Ser Phe Ala Pro Phe Ser Asn Pro Gly Asn Ala Glu Glu 865 870 875 880 Glu Cys Ala Thr Phe Ser Ala His Ser Gly Ser Leu Lys Lys Gin Ser 885 890 895 Pro Lys Val Thr Phe Glu Cys Glu Gin Lys Glu Glu Asn Gin Gly Lys 900 905 910 Asn Glu Ser Asn Ile Lys Pro Val Gin Thr Val Asn Ile Thr Ala Gly 915 920 925 Phe Pro Val Val Gly Gin Lys Asp Lys Pro Val Asp Asn Ala Lys Cys 930 935 940 Ser Ile Lys Gly Gly Ser Arg Phe Cys Leu Ser Ser Gln Phe Arg Gly 945 950 955 960 Asn Glu Thr Gly Leu Ile Thr Pro ADr' Lys His Gly Leu Leu Gin Asn 965 70 975 Pro Tyr Arg Ile Pro Pro Leu Phe Pro Ie Lys Ser Phe Val Lys Thr 980 985 990 Lys Cys Lys Lys Asn Leu Leu Glu Glu Asn Phe Glu Glu His Ser Met 995 1000 1005 Ser Pro Glu Arg Glu Met Gly Asn Glu Asn Ile Pro Ser Thr Val Ser 1010 1015 1020 Thr Ile Ser Arg Asn Asn Ile Arg Glu Asn Val Phe Lys Glu Ala Ser 1025 1030 1035 1040 Ser Ser Asn Ile Asn Glu Val Gly Ser Ser Thr Asn Glu Val Gly Ser 1045 1050 loss Ser Ile Asn Glu Ile Gly Ser Ser Asp Glu Asn Ile Gin Ala Glu Leu 1060 1065 1070 Gly Arg Asn Arg Gly Pro Lys Leu Asn Ala Met Leu Arg Leu Gly Val 1075 1080 1085 Leu Gin Pro Glu Val Tyr Lys Gin Ser Leu Pro Gly Ser Asn Cys Lys 1090 1095 1100 His Pro Glu Ile Lys Lys Gin Glu Tyr Glu Glu Val Val Gin Thr Val 1105 1110 1115 1120 L. 70 75 57

I

WO 96/33271 PCTIUS96/05621 Asn Thr Asp Phe Ser Pro Tyr Leu Ile Ser Asp Asn Leu Glu Gin Pro 1125 1130 1135 Met Gly Ser Ser His Ala Ser Gin Val Cys Ser Glu Thr Pro Asp Asp 1140 1145 1150 Leu Leu Asp Asp Gly Glu Ile Lys Glu Asp Thr Ser Phe Ala Glu Asn 1155 1160 1165 Asp Ile Lys Glu Ser Ser Ala Val Phe Ser Lys Ser Val Gin Lys Gly 1170 1175 1180 Glu Leu Ser Arg Ser Pro Ser Pro Phe Thr His Thr His Leu Ala Gin 1185 1190 1193 1200 Gly Tyr INFORMATION FOR SEQ ID NO:23: SEQUENCE CHARACTERISTICS: LENGTH: 1363 amino acids TYPE: amino acid ;.IlNDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gin Asn Val Ile Asn 1 5 10 Ala Met Gin Lys Ile Leu Glu Cys Pro Ile Cys Leu Glu Leu Ile Lys 20 25 Glu Pro Val Ser Thr Lys Cys Asp His Ile Phe Cys Lys Phe Cys Met 40 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Ser Gin Cys Pro Leu Cys 55 Lys Asn Asp Ile Thr Lys Arg Ser Leu Gin Glu Ser Thr Arg Phe Ser 70 75 Gin Leu Val Glu Glu Leu Leu Lys Ile Ile Cys Ala Phe Gin Leu Asp 90 Thr Gly Leu Glu Tyr Ala Asn Ser Tyr Asn Phe Ala Lys Lys Glu Asn 100 105 110 Asn Ser Pro Glu His Leu Lys Asp Glu Val Ser Ile Ile Gin Ser Met x 115 120 125 Gly Tyr Arg Asn Arg Ala Lys Arg Leu Leu Gin Ser Glu Pro Glu Asn 130 135 140 Pro Ser Leu Gln Glu Thr Ser Leu Ser Val Gin Leu Ser Asn Leu Gly 145 150 155 160 Thr Val Arg Thr Leu Arg Thr Lys Gin Arg Ile Gin Pro Gin Lys Thr 165 170 175 71

I'

f.Lb Z.Lw J x 260 265 58 WO 96/3327 1 Ser Val Lys Ala Pro Gin 210 Ala Cys 225 Pro Ser His Pro Pro Cys Ser Leu 290 Cys Asn 305 Trp Ala Glu Lys Trp Asn Asp Val 370 Trp Phe 385 Gly Glu Asn Giu Ala Ser Ser Lys 450 Tyr Arg 465 Leu Ile Giu Cys Arg Ser Asp 245 Tyr Asn Thr Lys Lys 325 Asp Lys Ile Ser Ser 405 Glu His Glu Ala Ala 485 Leu S er Asp Giu 230 Leu Gin Thr Lys Gin 310 Glu Leu Leu Thr Asp 390 Asn Tyr Glu Ser Ser 470 Phe PCTIUS96105621 Thr Vai Asn 190 Gin Ile Thr Lys Lys Ala His His Gin 240 Ala Giu Arg 255 His Val Glu 270 Giu Asn Ser Ala Glu Phe His Asn Arg 320 Pro Ser Thr 335 Arg Lys Glu 350 Asp Thr Glu Val Asn Giu Ser His Asp 400 Asp Val Leu 415 Asp Leu Leu 430 Arg Val His Gly Lys Thr Thr Glu Asn 480 Gin Glu Arg 495 Ser Gly Leu 510 Pro Leu Thr Asn Lys Leu Lys Arg 500 Pro Gin G.Ly Tnr arg A~sp ti.u .L.Ltemt- 210 215 220 WO 96/33271 His Pro Giu Asp Phe Ile Lys 515 PCTUS96/05621 Ala Asp Leu Ala Val Gin Lys Thr 525 Met PAsn Gin Ala Glu 595 Leu Va 1 Ser Pro Ala 675 Arg Gly Asn Lys Glu 755 Leu Glu Ser Ser His 835 Ile Ile Asn Phe 580 Leu Arg Ser Cys Val 660 Thr His Ser Pro Val1 740 Arg Val1 Val Gin Lys 820 Giu Gin Asn 550 Lys Thr Leu Lys Asn 630 Ser His Ala Ser Thr 710 Leu Asn Leu Gly Thr 790 Ala Asn Asn Giy Gly Giu 575 Ile Lys Leu Leu Tyr 655 Gly Gin Thr Lys Leu 735 Met S er Ile Asn Ile 815 Tyr Met Gin Asp 560 Lys Ser Lys Giu Gin 640 Asn Lys Thr Asn Giu 720 Giu Leu Ser Ser Lys 800 His Pro Glu va. o on flS %ZC y f2l± Z U &mLWX *UU.Jjjy 545 550 555 560 WO 96/33271 PCTIUS96/05621 Giu Ser Glu Leu Asp Ala Gln Tyr Leu Gln Asn Thr Phe Lys Val Ser 850 855 860 Lys Arg Gin Ser Phe Ala Pro Phe Ser Asn Pro Gly Asn Ala Glu Glu 865 870 875 880 Glu Cys Ala Thr Phe Ser Ala His Ser Gly Ser Leu Lys Lys Gln Ser 885 890 895 Pro Lys Val Thr Phe Glu Cys Glu Gln Lys Glu Glu Asn Gln Gly Lys 900 905 910 Asn Glu Ser Asn Ile Lys Pro Val Gln Thr Val Asn Ile Thr Ala Gly- 915 920 925 Phe Pro Val Val Gly Gin Lys Asp Lys Pro Val Asp Asn Ala Lys Cys 930 935 940 Ser Ile Lys Gly Gly Ser Arg Phe Cys Leu Ser Ser Gin Phe Arg Gly 945 950 955 960 Asn Glu Thr Gly Leu Ile Thr Pro Asn Lys His Gly Leu Leu Gin Asn 965 970 975 Pro Tyr Arg Ile Pro Pro Leu Phe Pro Ile Lys Ser Phe Val Lys Thr 980 985 990 Lys Cys Lys Lys Asn Leu Leu Glu Giu Asn Phe Giu Giu His Ser Met 995 1000 1005 Ser Pro Glu Arg Glu Met Gly Asn Glu Asn Ile Pro Ser Thr Val Ser 1010 1015 1020 Thr Ile Ser Arg Asn Asn Ile Arg Giu Asn Val Phe Lys Glu Ala Ser 1025 1030 1035 1040 Ser Ser Asn Ile Asn Giu Val Gly Ser Ser Thr Asn Giu Val Gly Ser 1045 1050 1055 Ser Ilie Asn Glu Ile Gly Ser Ser Asp Giu Asn Ile Gin Ala Giu Leu 1060 1065 1070 Gly Arg Asn Arg Gly Pro Lys Leu Asn Ala Met Leu Arg Leu Gly Val 1075 1080 1085 Leu Gin Pro Giu Val Tyr Lys Gin Ser Leu Pro Gly Ser Asn Cys Lys 1090 1095 1100 His Pro Glu Ile Lys Lys Gin Giu Tyr Giu Glu Val Val Gin Thr Val 1105 1110 1115 1120 Asn Thr Asp Phe Ser Pro Tyr Leu Ile Ser Asp Asn Leu Glu Gin Pro 1125 1130 1135 Met Gly Ser Ser His Ala Ser Gin Val Cys Ser Giu Thr Pro Asp Asp 1140 1145 1150 Leu Leu Asp Asp Gly Glu Ile Lys Giu Asp Thr Ser Phe Ala Giu Asn 1155 1160 1165 Asp Ile Lys Glu Ser Ser Ala Val Phe Ser Lys Ser Val Gin Lys Gly 1170 1175 1180 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Ser Gin Cys Pro Leu Cys 55 61 W 9633271 pCTIUS96/05621 Glu Leu Ser Arg Ser Pro Ser Pro Phe Thr His Thr His Leu Ala Gin 1185 1190 1195 1200 Gly Tyr Arg Arg Gly Ala Lys Lys Leu Glu Ser Ser Glu Glu Asn Leu 1205 1210 1215 Ser Ser Glu Asp Glu Glu Leu Pro Cys Phe Gin His Leu Leu Phe Gly 1220 1225 1230 Lys Val Asn Asn Ile Pro Ser Gln Ser Thr Arg His Ser Thr Val Ala 1235 1240 1245 Thr Glu Cys Leu Ser Lys Asn Thr Glu Glu Asn Leu Leu Ser Leu Lys 1250 1255 1260 Asn Ser Leu Asn Asp Cys Ser Asn Gin Val Ile Leu Ala Lys Ala Ser 1265 1270 1275 1280 Gin Glu His His Leu Ser Glu Glu Thr Lys Cys Ser Ala Ser Leu Phe 1285 1290 1295 Ser Ser Gin Cys Ser Glu Leu Glu Asp Leu Thr Ala Asn Thr Asn Thr 1300 1305 1310 Gin Asp Pro Phe Leu Ile Gly Ser Ser Lys Gin Met Arg His Gin Ser 1315 1320 1325 Glu Ser Gin Gly Val Gly Leu Ser Asp Lys Glu Leu Val Ser Asp Asp 1330 1335 1340 Glu Glu Arg Gly Thr Gly Leu Glu Glu Asn Lys Lys Ser Lys Ala Trp 1345 1350 1355 1360 Ile Gin Thr INFORMATION FOR SEQ ID NO:24: SEQUENCE CHARACTERISTICS: LENGTH: 1852 amino acids TYPE. amino acid STRAIKDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: Met Asp Leu Ser Ala Leu Arg Val Glu Glu Val Gin Asn Val Ile Asn 1 5 10 Ala Met Gin Lys Ile Leu Glu Cys Pro Ile Cys Leu Glu Lou Ile Lys 25 Giu Pro Val Ser Thr Lys Cys Asp His Ile Phe Cys Lys Phe Cys Met 40 Leu Lys Leu Leu Asn Gin Lys Lys Gly Pro Ser Gin Cys Pro Leu Cys 55 Lys Asn Asp Ile Thr Lys Arg Ser Leu Gin Glu Ser Thr Arg Phe Ser 70 75 Trp Phe Ser Arg Ser Asp Glu Leu Leu Gly Ser Asp Asp Ser His Asp 385 390 395 400 WO 96/33271 Gin Leu Thr Gly Asn Ser Gly Tyr 130 Pro Ser 145 Thr Val Ser Val Lys Ala Pro Gin 210 Ala Cys 225 Pro Ser Hius Pro Pro Cys Ser Leu 290 Cys Asn 305 Trp Ala Giu Lys Trp Asn Asp Val 370 Trp Phe 385 Gly Giu PCT/US96/05621l Gin Leu Asp Lys Giu Asn 110 Gin Ser Met Pro Giu Asn Asn Leu Gly 160 Gin Lys Thr 175 Thr Val Asn 190 GinIlie Thr Lys Lys Ala H-is His Gin 240 Ala Giu Arg 255 His Val Giu 270 Giu Asn Ser i±a Giu Phe s Asn An Pro Ser Thr Arg Lys Giu '150 Thr C Val Asii Giu Ser His Asp 400 Asp Val Leu 415 WO 96/3,

A.,

Se Ty 46 Lei Pr( HiE, Prc Val 545 Ser Giu Asn Asn Leu.

625 Ilie Gin Glu Ser Ala 705 Phe V Thr v '1 4 PheVa Afl roS~ Leu Pro Arg Glu Glu LYS GiU UluU Lyb 1 ~7257375 WO 96/3327 1 Asn Glu Val Asp Glu Tyr Ser Gly Ser Ser Glu Lys Ile 420 425 PCTIUS96/05621 Asp Leu Leu 430 Ala Ser Tyr 465 Leu Pro His Pro Val 545 Ser Glu Asn Asn Leu 625 Ile Gin Glu Ser Al a 705 Phe Thr Asp 435 S er Lys Ile Thr Glu 515 Met Asn Gin Al a Glu 595 Leu Val1 Ser Pro Al a 675 Arg Gly Asn Lys Pro Val Lys Gly Asn 500 Asp Ile Ile Asn Phe 580 Leu Arg Ser Cys Val 660 Thr His S er Pro Val 740 Ile Glu Asn Giu Lys 505 Ala Asn His Asn Glu 585 His Thr Pro Glu Asn 665 Ser Phe S er Glu Glu 745 Arg Gly Thr Gin Ser 510 Gin Asn Lys Leu Ser 590 Pro Ala Giu Lys Giu 670 Giu Leu Leu Lys Leti 750 Val Lys Gilu Gi u 495 Gly Lys Gly Gly Giu 575 Ile Lys Leu Leu Tyr 655 Gly Gin Thr Lys Leu 735 Met His Thr Asn 480 Arg Leu Thr Gin Asp 560 Lys S er Lys Glu Gin 640 Asn Lys Thr Asn Glu 720 Glu Leu Gin Leu Val Giu Giu Leu Leu Lys Ile Ile Cys Ala Phe Gin Leu Asp 90 WO 96/33271 PCTUJS96/05621 Ser Gly Giu Arg Val Leu Gin Thr Giu Arg Ser Val Giu Ser Ser Ser 755 760 765 Ile Ser Leu Vai 770 Leu Leu Giu Val 785 Cys Val Ser Gin Gly Cys Ser Lys 820 Leu Giy His Giu 835 Giu Ser Giu Leu 850 Lys Arg Gin Ser 865 Giu Cys Ala Thr Pro LyG Vai Thr 900 Asn Giu Ser Asn 915 Phe Pro Vai Val 930 Ser Ile Lys Gly 945 Asn Giu Thr Gly Pro Tyr Arg Ile 980 Lys Cys Lys Lys 995 Ser Pro GJlu Arg 1010 Thr Ile Ser Arg 1025 Ser Ser Asn Ile Pro Gly Thr Asp Tyr 775 Ser Thr Leu Gly Lys 790 Cys Ala Ala Phe Giu 805 Asp Asn Arg Asn Asp 825 Val Asn His Ser Arg 840 Asp Ala Gin Tyr Leu 855 Phe Ala Pro Phe Ser 870 Phe Ser Ala His Ser 885 Phe Giu Cys Giu Gin 905 Ile Lys Pro Vai Gin 920 Giy Gin Lys Asp Lys 935 Giy Ser Arg Phe Cys 950 Leu Ile Thr Pro Asin 965 Pro Pro Leu Phe Pro 985 Asn Leu Leu Giu Giu 1000 Giu Met Gly Asn Glu 10i5 Asn Asn Ile Arg Giu 1030 Asn Giu Vai Giy Ser 1045 Ile Giy Ser Ser Asp 0 1065 Gly Thr Gin Giu Ser Ile Ser 780 Ala Lys Thr Giu Pro Asn Lys 795 800 Asn Pro Lys Gly Leu Ile His 810 815 Thr Giu Giy Phe Lys Tyr Pro.

830 Giu Thr Ser Ile Glu Met Giu 845 Gin Asn Thr Phe Lys Vai Ser 860 Asn Pro Gly Asn Aia Giu Giu 875 880 Giy Ser Leu Lys Lys Gin Ser 890 895 Lys Giu Giu Asn Gin Giy Lys 910 Thr Vai Asn Ile Thr Aia Gly 925 Pro Val Asp Asn Ala Lys Cys 940 Leu Ser Ser Gin Phe Arg Giy 955 960 Lys His Giy Leu Leu Gin Asn 970 975 Ile Lys Ser Phe Vai Lys Thr 990 Asn Phe Giu Giu His Ser Met i005 Asn Ile Pro Ser Thr Vai Ser 1020 Asn Vai Phe Lys Giu Aia Ser 1035 1040 Ser Thr Asn Giu Vai Giy Ser 1050 1055 Giu Asn Ile Gin Ala Giu Leu 1070 Ala Met Leu Arg Leu Giy Val 1085 Ser Ile Asn Giu 106( Gly Arg Asn 1075 Arg Gly Pro Lys Leu Asn 1080 Asn Glu Val Asp Glu Tyr Ser Gly Ser Ser Glu Lys Ile Asp Leu Leu 420 425 430 51

B!

WO 96/33271 PCT/US96/05621 Leu Gin Pro Glu Val Tyr Lys Gin Ser Leu Pro Gly Ser Asn Cys Lys 1090 1095 1100 His Pro Glu Ile Lys Lys Gin Glu Tyr Glu Glu Val Val Gin Thr Val 1105 1110 1115 1120 Asn Thr Asp Phe Ser Pro Tyr Leu Ile Ser Asp Asn Leu Glu Gin Pro 1125 1130 1135 Met Gly Ser Ser His Ala Ser Gin Val Cys Ser Glu Thr Pro Asp Asp 1140 1145 1150 Leu Leu Asp Asp Gly Glu Ile Lys Glu Asp Thr Ser Phe Ala Glu Asn 1155 1160 1165 Asp Ile Lys Glu Ser Ser Ala Val Phe Ser Lys Ser Val Gin Lys Gly 1170 1175 1180 Glu Leu Ser Arg Ser Pro Ser Pro Phe Thr His Thr His Leu Ala Gin 1185 1190 1195 1200 Gly Tyr Arg Arg Gly Ala Lys Lys Leu Glu Ser Ser Glu Glu Asn Leu 1205 1210 1215 Ser Ser Glu Asp Glu Glu Leu Pro Cys Phe Gin His Leu Leu Phe Gly 1220 1225 1230 Lys Val Asn Asn Ile Pro Ser Gin Ser Thr Arg His Ser Thr Val Ala 1235 1240 1245 Thr Glu Cys Leu Ser Lys Asn Thr Glu Glu Asn Leu Leu Ser Leu Lys 1250 1255 1260 Asn Ser Leu Asn Asp Cys Ser Asn Gin Val Ile Leu Ala Lys Ala Ser 1265 1270 1275 1280 Gin Glu His His Leu Ser Glu Glu Thr Lys Cys Ser Ala Ser Leu Phe 1285 1290 1295 Ser Ser Gin Cys Ser Glu Leu Glu Asp Leu Thr Ala Asn Thr Asn Thr 1300 1305 1310 Gin Asp Pro Phe Leu Ile Gly Ser Ser Lys Gin Met Arg His Gln Ser 1315 1320 1325 Glu Ser Gin Gly Val Gly Leu Ser Asp Lys Glu Leu Val Ser Asp Asp 1330 1335 1340 Glu Glu Arg Gly Thr Gly Leu Glu Glu Asn Asn Gin Glu Glu Gin Ser 1345 1350 1355 1360 Met Asp Ser Asn Leu Gly Glu Ala Ala Ser Gly Cys Glu Ser Glu Thr 1365 1370 1375 Ser Val Ser Glu Asp Cys Ser Gly Leu Ser Ser Gin Ser Asp Ile Leu 1380 1385 1390 Thr Thr Gin Gin Arg Asp Thr Met Gin His Asn Leu Ile Lys Leu Gin 1395 1400 1405 Gin Glu Met Ala Glu Leu Glu Ala Val Leu Glu Gin His Gly Ser Gin 1410 1415 1420 I 79 i WO 96/33271 PCTIUS96/05621 Pro Ser Asn Ser Tyr Pro Ser Ile Ile Ser Asp Ser Ser Ala Leu Giu 1425 1430 1435 1440 Asp Leu Arg Asn Pro Giu Gin Ser Thr Ser Giu Lys Ala Val Leu Thr 1445 1450 1455 Ser Gln Lys Ser Ser Giu Tyr Pro Ile Ser Gln Asn Pro Giu Gly Leu 1460 1465 1470 Ser Ala Asp Lys Phe Glu Val Ser Ala Asp Ser Ser Thr Ser Lys Asn 1475 1480 1485 Lys Giu Pro Gly Vai Giu Arg Ser Ser Pro Ser Lys Cys Pro Ser Leu 1490 1495 1500 Asp Asp Arg Trp Tyr Met His Ser Cys Ser Gly Ser Leu Gin Asn Arg 1505 1510 1515 1520 Asn Tyr Pro Ser Gin Glu Giu Leu Ile Lys Val Val Asp Vai Glu Gl-i 1525 1530 1535 Gin Gin Leu Giu Giu Ser Gly Pro His Asp Leu Thr Giu Thr Ser Tyr 1540 1545 1550 L~u Pro Arg Gin Asp Leu Giu Gly Thr Pro Tyr Leu Giu Ser Gly Ile 1555 1560 1565 Ser Leu Phe Ser Asp Asp Pro Giu Ser Asp Pro Ser Giu Asp Arg Ala 1570 1575 1580 Pro Glu Ser Ala Arg Val Gly Asn lie Pro ber Ser Thr Ser Ala Leu 1585 1590 1595 1600 Lys Val Pro Gin Leu Lys Val Ala Giu Ser Ala Gin Ser Pro Ala Ala 1605 1610 1615 Ala His Thr Thr Asp Thr Ala Gly Tyr Asn Ala Met Giu Glu Ser Val 1620 1625 1630 Ser Arg Giu Lys Pro Giu Leu Thr Ala Ser Thr Giu Arg Val Asn Lys 1635 1640 1645 Arg Met Ser Met Val Val Ser Gly Leu Thr Pro Giu Glu Phe Met Leu 1650 1655 1660 Val Tyr Lys Phe Ala Arg Lys His His Ile Thr Leu Thr Asn Leu Ile 1665 1670 1675 1680 Thr Giu Giu Thr Thr His Val Vai Met Lys Thr Asp Aia Giu Phe Val 1685 1690 1 395 Cys Giu Arg Thr Leu Lys Tyr Phe Leu Giy Ile Ala Giy Gly Lys Trp 1700 1705 1710 Val Val Ser Tyr Phe Trp Val Thr Gin Ser Ile Lys Giu Arg Lys Met 1715 1720 1725 Le~u Asn Giu His Asp Phe Giu Val Arg Giy Asp Val Val Asn Giy Arg 1730 1735 1740 Asn His Gin Gly Pro Lys Arg Ala Arg Giu Ser Gin Asp Arg Lys Ile 1745 1750 1755 1760 WO 96/33271 PCTIUS96/05621 Phe Arg Gly Leu Giu Ile Cys Cys Tyr GlY Pro Phe Thr Asn Met Pro 1765 1770 1775 Thr Asp Gin Leu Giu Trp Met Val Gin Leu Cys Giy Ala Ser Vai Val 51780 1785 1790 Lys Giu Leu Ser Ser Phe Thr Leu Gly Thr Giy Vai His Pro Ile Val 1795 1800 1805 Vai Val Gin Pro Asp Ala Trp Thr Glu Asp Asn Gly Phe His Ala Ile 1810 1815 1820 Gly Gin Met Cys Giu Ala Pro Val Val Thr Arg Giu Trp Val Leu Asp 1825 1830 1835 1840 Ser Val Ala Leu Tyr Gin Cys Gin Giu Leu Asp Thr 1845 IRC;0 81 A9

Claims (12)

1. isolated nucleic acid comprising BRCAI allele 95803 (SEQ ED NO:1t), 9601 (SEQ ID 9815 (SEQ ID 1NO1:3),82203 (SEQ ID NO:5), 388 (SEQUENCE ED NO:6), or a fragment thereof, wherein said fragment is capable of specifically h-ybridizin~g with said allele in the presence of wld-type 13RCAI.

2. An isolated nucleic acid comprising BRCAl allele #5803 (SEQ ID NO: 1) or a fragment thereof, wherein said fragment is capable of specifically hybridizing with said allele in the presence of wild-type BRCAI.

3. An isolated nucleic acid comprising )3RCA. allele "13601 (SEQ ED NO:2) or a fragment thereof, wherein said fragment is capable of specifically hybridizing with said allele in the presence of wild-type BRCAl.

4. An isolated nucleic acid comprising BRCAI allele M981.5 (SE-Q ED NO:3) or a frag,?ment thereof, wherein said fragment is capable of specifically hybridizing with said allele in the presence of wild-type BIRCAI. An isolated nucleic acid comprising BRCAI allele '#t8203 (SEQ ID NO-5') of a fragment thereof, wherein said figmni 13 i capable of specifically hybridizing with said allele in th~e presence of wild-type BRCA1.

6. An isolated nucleic acid comprising BRCA1 allele #388 (SEQUENCE ID NO:6), or a fragment thereof, wherein said fragment is capable of specifically hybridizing witlh said allele in the presence of wild-type BRCA1.

7. An isolated translation product of BRCAI allel e #6 803 (SEQ ID_ NO: t13), 96 01 (SEQ ID NO: 14), 9815 (SEQ ED NO: 15), 8203 (SEQ ED NO: 17), 388 (SEQ ID _NO: 18), or a C-terminus fragment thereof

8. An isolated translation product of BRCA1 allele 803 (SEQ ED NO- 13) or a C- terminus fragment thereof ~NrO~AMENDED SHEET TI

9. An isolated translation product ofBRCA1 alleLe #9601 (SEQ D NO: 14) or a C- terminus fragment thereof. An isolated translation product ofBRCA1 allele #98 5 (SEQ ID NO:15) or a C- terminus fragment thereof.

11. An isolated translation product of BRCA1 allele #8203 (SEQ ID NO:17) or a C- terminus fragment thereof.

12. An isolated translation product of BRCA1 allele 388 (SEQ ID NO:18) or a C- terminus fragment thereof.

13. A method of screening a patient for a cancer a.e susceptibility, said method comprising the steps of: isolating from said patient a first nucleic acid comprising e0* at least one BRCA1 allele or fragment thereof; contacting said sample with a second nucleic acid according to any one of claims 1, 2, 3, 4, 5 and 6, under conditions whereby said second nucleic acid is capable of specifically hybridizing with said first nucleic acid; detecting the presence or absence of specific hybridization of said second nucleic acid with said first nucleic acid; wherein the presence of specific hybridization of said second nucleic acid to said first nucleic acid is diagnostic of a cancer susceptibility. e e e2

14. A method of screening a patient for a cancer susceptibility, said method comprising the steps of: isolating from said patient a composition comprising a first translation product of at least one BRCA1 allele; contacting said first translation product with a BRCA1 gene product specific binding agent specific for a protein or C-terminal fragment thereof according to any one of claims 7, 8, 9, 10, 11 and 12 under conditions wherein said reagent is capable of specifically binding said first translation product; detecting the presence or absence of specifically bound complexes of said reagent and said first translation product; wherein the presence of said complexes correlates with a cancer susceptibility.